Lavage and/or infusion using CSA compounds for increasing fertility in a mammal

ABSTRACT

Increasing fertility in a mammal utilizes cationic steroidal antimicrobial (CSA) compounds and CSA-containing compositions. Such treatment in a mammal includes administering a formulation (e.g., lavage and/or infusion) including at least one CSA compound, or pharmaceutically acceptable salt thereof, to the reproductive structure(s) of the mammal (e.g., horse, dairy cow, human, etc.). The formulation may be applied topically as lavage and/or infusion to desired reproductive structures, such as the vagina, cervix, uterus, penis, or combinations thereof. The formulation may kill both planktonic and biofilm forms of sperm killing microbes, and may at least partially break up a microbial plaque or film located within any of the reproductive structures (e.g., the uterus). The CSA or pharmaceutically acceptable salt thereof may be selective or preferential in its action, so as to preferentially kill sperm killing microbes without causing harm to beneficial microbes also residing within the reproductive structure of the mammal.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.61/952,682 filed Mar. 13, 2014, the disclosure of which is incorporatedherein in its entirety.

BACKGROUND OF THE INVENTION

1. The Field of the Invention

The present invention relates to compositions and methods for increasingfertility in a mammal using cationic steroidal antimicrobial (CSA)compounds.

2. The Relevant Technology

Maintaining fertility in dairy cows, horses, domestic animals, and othermammals continues to be of importance to those engaged in the farmingindustry. Loss of fertility greatly reduces the value of breedinganimals, which can only then be used for meat but which may not besuitable for such use. Thus, fertility loss, especially prematureinfertility, may substantially or completely destroy the economic valueof an animal relied upon for breeding.

In general, there may be many reasons to restore or enhance fertility ina wide variety of different mammals, including humans. Currenttreatments to enhance fertility include administration of fertilitydrugs, including hormones or other substances that act by acceleratingovulation, and/or surgical procedures, some of which can be veryinvasive and expensive.

In addition to old age, infertility may also be caused by fungal orbacterial endometritis, which occurs when a bacteria or fungus (e.g.,yeast) contaminates the uterus and causes infection and inflammation. Insome cases, endometritis can be sexually transmitted, such as ContagiousEquine Metritis (CEM), caused by Taylorella equigenitalis, a Gramnegative coccobacillus. The bacteria can survive for an extended periodon the external genitalia of the stallion and the vagina or clitoris ofthe mare. When infected, the stallion may show no clinical signs. Themare, however, typically presents acute endometritis with a thick, grey,mucoid discharge after breeding and may short cycle due to inflammation.The acute signs may subside rapidly, with some mares remaining asasymptomatic carriers.

Endometritis is difficult to treat and has a high rate of recurrence.Current treatments may include uterine lavage with large volumes offluid and/or administration of dimethylsulfoxide (DMSO), antifungalagents (e.g., clotrimazole, amphotericin, fluconazole, nystatin), orantibiotics. Such treatments are expensive and outcomes uncertain. Theprognosis is often poor since treatment only affects certain stages ofbacterial or fungal development. In some cases, the microbe may beattached within folds of the endometrium, or the mare may have delayeduterine clearance. Even with successful treatment, chance ofre-infection is high.

Accordingly, there remains a present but unsatisfied need to findreliable treatments for increasing fertility in mammals.

SUMMARY

Disclosed herein are compositions and methods for enhancing fertility ina mammal. According some embodiments, one or more CSA compounds areadministered to the reproductive structure (e.g., uterus or penis) of amammal in order to increase fertility, such as in the form of a lavageand/or infusion. The present methods and use of the compositionsdisclosed herein have surprisingly been shown to allow previouslyinfertile mammals to conceive, including mammals which did not respondto and become fertile using conventional lavage or other fertilitytreatments.

For reasons that may not be entirely understood, administeringCSA-containing compositions to the uterus or other reproductivestructure of a female mammal in the form of a lavage and/or infusionhave unexpectedly been found to increase fertility and cause conceptionin a mammal that was previously unable to conceive. It is also believedthat topical application of CSA-containing compositions to the penis ofa male mammal prior to copulation may also enhance fertility of therecipient female mammal. According to one theory, the CSA-containingcompositions treat infertility by breaking up microbial plaque or filmlocated in a uterus (e.g., metritis or endometritis). It is believedthat the CSA-containing composition hastens uterine epithelial cellhealing (due to its epitheliotrophic activity) and modulates theinflammation (due to enhanced innate immune response).

For example, infertility may be caused by fungal or bacterialendometritis, which occurs when a bacteria, fungus (e.g., yeast), etc.contaminates the uterus and causes infection and inflammation. Thisproblem is difficult to treat, and has a high rate of recurrence.Current treatments may include uterine lavage with large volumes offluid or administration of dimethylsulfoxide (DMSO), antifungal agents(e.g., clotrimazole, amphotericin, fluconazole, nystatin), orantibiotics. Such treatments are expensive and outcomes uncertain as theprognosis is often poor since treatment only affects certain stages ofbacterial or fungal development and/or the organism may be attachedwithin folds of the endometrium, or the mare may have delayed uterineclearance. Even with successful treatment, chance of re-infection ishigh. The present methods and use of the compositions disclosed hereinhave surprisingly been shown to allow mammals to conceive, where themammals were previously infertile and/or did not positively respond toconventional fertility treatments.

As part of the treatment process, CSA-containing compositions may beformulated to increase efficacy and/or reduce cytotoxicity to mammals.In some embodiments, the CSA-containing compositions can be formulatedin order to deliver CSA compounds as individually dispersed CSAmolecules and/or very small CSA particles having low agglomeration. Inone embodiment, CSA-containing compositions are formulated in order toremain stable and resist agglomeration of CSA molecules or particles tolarger CSA particles over a specified time period. This may beaccomplished, for example, by the use of micelle-forming agents whichare able to form micelles in a liquid carrier, which encapsulate the CSAmolecules and prevent or reduce agglomeration.

Additional features and advantages will be set forth in part in thedescription that follows, and in part will be obvious from thedescription, or may be learned by practice of the embodiments disclosedherein. The objects and advantages of the embodiments disclosed hereinwill be realized and attained by means of the elements and combinationsparticularly pointed out in the appended claims. It is to be understoodthat both the foregoing brief summary and the following detaileddescription are exemplary and explanatory only and are not restrictiveof the embodiments disclosed herein or as claimed.

DETAILED DESCRIPTION

Disclosed herein are methods and systems for applying CSA-containingcompositions to the genitalia or other reproductive structure of amammal to enhance fertility. According to some embodiments, aCSA-containing composition is administered to the uterus or otherreproductive structure of a female mammal in order to increasefertility, such as in the form of a lavage and/or infusion. According toother embodiments, a CSA-containing composition is applied to the penisof a male mammal prior to copulation with a female mammal. In someembodiments, the CSA-containing compositions may be formulated tocontrol or reduce CSA agglomeration, increase stability, enhanceefficacy, and/or reduce cytotoxicity to mammalian cells.

For reasons that may not be entirely understood, administering aCSA-containing composition to the uterus or other reproductive structureof a mammal has unexpectedly been found to increase fertility and causeconception in a mammal that was previously unable to conceive. It isalso believed that topical application of a CSA-containing compositionto the penis of a male mammal prior to copulation may also enhancefertility of the recipient female mammal.

According to one theory, CSA-containing compositions act by breaking upbiofilms and/or killing microbes that form biofilms present in theuterus or other reproductive structure of a mammal that might inhibitfertilization. Such biofilms may form a barrier between gametes (i.e.,sperm cells and the ovum) and/or prevent implantation of a fertilizedzygote within the uterus and/or cause spontaneous abortion of a zygoteshortly after implantation. It is believed that CSA-containingcompositions are able to break up biofilms and/or kill microbes thatform such biofilms as a result of the unique amphiphilic nature andchemical functionalities of CSA compounds. The CSA-containingcomposition can also accelerate uterine epithelial cell healing due toits epitheliotrophic activity and modulates the inflammation due toenhanced innate immune response.

As part of the treatment process, CSA-containing compositions may beformulated to increase efficacy and/or reduce cytotoxicity to mammals.In some embodiments, the CSA-containing compositions can be formulatedto deliver CSA compounds as individually dispersed CSA molecules and/orvery small CSA particles having low agglomeration. In one embodiment,CSA-containing compositions are formulated in order to remain stable andresist agglomeration of CSA molecules or CSA particles to larger CSAparticles over a predetermined time period. This may be accomplished,for example, by the use of micelle-forming agents that are able to formmicelles in a liquid carrier, which encapsulate the CSA molecules andprevent or reduce agglomeration. Additional details regardingcompositions and methods for forming stable CSA-containing compositionswith reduced agglomeration are disclosed in U.S. Provisional ApplicationNo. 61/952,669, filed Mar. 13, 2014, the disclosure of which isincorporated herein in its entirety.

Before describing exemplary compositions and methods for forming CSAcompositions for treating infertility, as well as a more detaileddescription of properties, chemical characteristics, and uses, a generaldescription will be given of non-limiting examples of CSA compounds thatcan be used in the disclosed CSA compositions and methods.

I. CSA COMPOUNDS AND COMPOSITIONS

Cationic steroidal anti-microbial (CSA) compounds, sometimes referred toas “CSAs” or “ceragenin compounds”, can include synthetically produced,small molecule chemical compounds that include a sterol backbone havingvarious charged groups (e.g., amine and cationic groups) attached to thebackbone. The sterol backbone can be used to orient the amine orguanidine groups on one face, or plane, of the sterol backbone. CSAcompounds are cationic and amphiphilic, based upon the functional groupsattached to the backbone. They are facially amphiphilic with ahydrophobic face and a polycationic face. Without wishing to be bound toa particular theory, the CSA compounds described herein act asanti-microbial agents (e.g., anti-bacterials, antifungals, andanti-virals). It is believed, for example, that the CSA compounds act asanti-microbial agents by binding to the cellular membrane of bacteriaand other microbes and inserting into the cell membrane, forming one ormore pores, which allows leakage of ions and/or cytoplasmic materialscritical to the microbe's survival, leading to the death of the affectedmicrobe. In addition, the CSA compounds described herein may also act tosensitize bacteria to other antibiotics. For example, at concentrationsbelow the corresponding minimum bacteriostatic concentration for the CSAcompound(s), the CSA compound(s) may nevertheless cause bacteria tobecome more susceptible to other antibiotics by increasing membranepermeability. It is postulated that ionically charged groups areresponsible for disrupting the bacterial cellular membrane and impartinganti-microbial properties.

A. Definitions

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which these embodiments belong. The terminology used in thedescription herein is for describing particular embodiments only and isnot intended to be limiting of the embodiments. As used in thespecification and the appended claims, the singular forms “a,” “an,” and“the” are intended to include the plural forms as well, unless thecontext clearly indicates otherwise. All publications, patentapplications, patents, and other references mentioned herein areincorporated by reference in their entirety.

Terms and phrases used in this application, and variations thereof,especially in the appended claims, unless otherwise expressly stated,should be construed as open ended as opposed to limiting. As examples ofthe foregoing, the term “including” should be read to mean “including,without limitation,” “including but not limited to,” or the like; theterm “comprising” as used herein is synonymous with “including,”“containing,” or “characterized by,” and is inclusive or open-ended anddoes not exclude additional, unrecited elements or method steps; theterm “having” should be interpreted as “having at least”; the term“includes” should be interpreted as “includes but is not limited to”;the term “example” is used to provide exemplary instances of the item indiscussion, not an exhaustive or limiting list thereof; and use of termslike “preferably,” “preferred,” “desired,” or “desirable,” and words ofsimilar meaning should not be understood as implying that certainfeatures are critical, essential, or even important to the structure orfunction of the invention, but instead as merely intended to highlightalternative or additional features that may or may not be utilized in aparticular embodiment. In addition, the term “comprising” is to beinterpreted synonymously with the phrases “having at least” or“including at least”. When used in the context of a process, the term“comprising” means that the process includes at least the recited steps,but may include additional steps. When used in the context of acompound, composition or device, the term “comprising” means that thecompound, composition or device includes at least the recited featuresor components, but may also include additional features or components.Likewise, a group of items linked with the conjunction “and” should notbe read as requiring that each and every one of those items be presentin the grouping, but rather should be read as “and/or” unless expresslystated otherwise. Similarly, a group of items linked with theconjunction “or” should not be read as requiring mutual exclusivityamong that group, but rather should be read as “and/or” unless expresslystated otherwise.

It is understood that, in any compound described herein having one ormore chiral centers, if an absolute stereochemistry is not expresslyindicated, then each center may independently be of R-configuration orS-configuration or a mixture thereof. Thus, the compounds providedherein may be enantiomerically pure, enantiomerically enriched, racemicmixture, diastereomerically pure, diastereomerically enriched, or astereoisomeric mixture. In addition it is understood that, in anycompound described herein having one or more double bond(s) generatinggeometrical isomers that can be defined as E or Z, each double bond mayindependently be E or Z a mixture thereof.

Likewise, it is understood that, in any compound described, alltautomeric forms are also intended to be included.

It is to be understood that where compounds disclosed herein haveunfilled valencies, then the valencies are to be filled with hydrogensor isotopes thereof, e.g., hydrogen-1 (protium) and hydrogen-2(deuterium).

It is understood that the compounds described herein can be labeledisotopically. Substitution with isotopes such as deuterium may affordcertain therapeutic advantages resulting from greater metabolicstability, such as, for example, increased in vivo half-life or reduceddosage requirements. Each chemical element as represented in a compoundstructure may include any isotope of said element. For example, in acompound structure a hydrogen atom may be explicitly disclosed orunderstood to be present in the compound. At any position of thecompound that a hydrogen atom may be present, the hydrogen atom can beany isotope of hydrogen, including but not limited to hydrogen-1(protium) and hydrogen-2 (deuterium). Thus, reference herein to acompound encompasses all potential isotopic forms unless the contextclearly dictates otherwise.

It is understood that the methods and combinations described hereininclude crystalline forms (also known as polymorphs, which include thedifferent crystal packing arrangements of the same elemental compositionof a compound), amorphous phases, salts, solvates, and hydrates. In someembodiments, the compounds described herein exist in solvated forms withpharmaceutically acceptable solvents such as water, ethanol, or thelike. In other embodiments, the compounds described herein exist inunsolvated form. Solvates contain either stoichiometric ornon-stoichiometric amounts of a solvent, and may be formed during theprocess of crystallization with pharmaceutically acceptable solventssuch as water, ethanol, or the like. Hydrates are formed when thesolvent is water, or alcoholates are formed when the solvent is alcohol.In addition, the compounds provided herein can exist in unsolvated aswell as solvated forms. In general, the solvated forms are consideredequivalent to the unsolvated forms for the purposes of the compounds andmethods provided herein.

Unless otherwise indicated, all numbers expressing quantities ofingredients, reaction conditions, and so forth used in the specificationand claims are to be understood as being modified in all instances bythe term “about.” Accordingly, unless indicated to the contrary, thenumerical parameters set forth in the specification and attached claimsare approximations that may vary depending upon the desired propertiessought to be obtained by the present embodiments. At the very least, andnot as an attempt to limit the application of the doctrine ofequivalents to the scope of the claims, each numerical parameter shouldbe construed in light of the number of significant digits and ordinaryrounding approaches.

Notwithstanding that the numerical ranges and parameters setting forththe broad scope of the embodiments are approximations, the numericalvalues set forth in the specific examples are reported as precisely aspossible. Any numerical value, however, inherently contains certainerrors necessarily resulting from the standard deviation found in theirrespective testing measurements. Every numerical range given throughoutthis specification and claims will include every narrower numericalrange that falls within such broader numerical range, as if suchnarrower numerical ranges were all expressly written herein. Where arange of values is provided, it is understood that the upper and lowerlimit, and each intervening value between the upper and lower limit ofthe range is encompassed within the embodiments.

As used herein, any “R” group(s) such as, without limitation, R₁, R₂,R₃, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀, R₁₁, R₁₂, R₁₃, R₁₄, R₁₅, R₁₆, R₁₇, andR₁₈ represent substituents that can be attached to the indicated atom.Unless otherwise specified, an R group may be substituted orunsubstituted.

The term “ring” as used herein can be heterocyclic or carbocyclic. Theterm “saturated” used herein refers to a fused ring having each atom inthe fused ring either hydrogenated or substituted such that the valencyof each atom is filled. The term “unsaturated” used herein refers to afused ring where the valency of each atom of the fused ring may not befilled with hydrogen or other substituents. For example, adjacent carbonatoms in the fused ring can be doubly bound to each other. Unsaturationcan also include deleting at least one of the following pairs andcompleting the valence of the ring carbon atoms at these deletedpositions with a double bond; such as R₅ and R₉; R₈ and R₁₀; and R₁₃ andR₁₄.

Whenever a group is described as being “substituted” that group may besubstituted with one, two, three or more of the indicated substituents,which may be the same or different, each replacing a hydrogen atom. Ifno substituents are indicated, it is meant that the indicated“substituted” group may be substituted with one or more group(s)individually and independently selected from alkyl, alkenyl, alkynyl,cycloalkyl, cycloalkenyl, cycloalkynyl, acylalkyl, alkoxyalkyl,aminoalkyl, amino acid, aryl, heteroaryl, heteroalicyclyl, aralkyl,heteroaralkyl, (heteroalicyclyl)alkyl, hydroxy, protected hydroxyl,alkoxy, aryloxy, acyl, mercapto, alkylthio, arylthio, cyano, halogen(e.g., F, Cl, Br, and I), thiocarbonyl, O-carbamyl, N-carbamyl,O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, S-sulfonamido,N-sulfonamido, C-carboxy, protected C-carboxy, O-carboxy, isocyanato,thiocyanato, isothiocyanato, nitro, oxo, silyl, sulfenyl, sulfinyl,sulfonyl, haloalkyl, haloalkoxy, trihalomethanesulfonyl,trihalomethanesulfonamido, an amino, a mono-substituted amino group anda di-substituted amino group, R_(a)O(CH₂)_(m)O—, R_(b)(CH₂)_(n)O—,R_(c)C(O)O(CH₂)_(p)O—, and protected derivatives thereof. Thesubstituent may be attached to the group at more than one attachmentpoint. For example, an aryl group may be substituted with a heteroarylgroup at two attachment points to form a fused multicyclic aromatic ringsystem. Biphenyl and naphthalene are two examples of an aryl group thatis substituted with a second aryl group.

As used herein, “C_(a)” or “C_(a) to C_(b)” in which “a” and “b” areintegers refer to the number of carbon atoms in an alkyl, alkenyl oralkynyl group, or the number of carbon atoms in the ring of acycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl orheteroalicyclyl group. That is, the alkyl, alkenyl, alkynyl, ring of thecycloalkyl, ring of the cycloalkenyl, ring of the cycloalkynyl, ring ofthe aryl, ring of the heteroaryl or ring of the heteroalicyclyl cancontain from “a” to “b”, inclusive, carbon atoms. Thus, for example, a“C₁ to C₄ alkyl” group refers to all alkyl groups having from 1 to 4carbons, that is, CH₃—, CH₃CH₂—, CH₃CH₂CH₂—, (CH₃)₂CH—, CH₃CH₂CH₂CH₂—,CH₃CH₂CH(CH₃)— and (CH₃)₃C—. If no “a” and “b” are designated withregard to an alkyl, alkenyl, alkynyl, cycloalkyl cycloalkenyl,cycloalkynyl, aryl, heteroaryl or heteroalicyclyl group, the broadestrange described in these definitions is to be assumed.

As used herein, “alkyl” refers to a straight or branched hydrocarbonchain that comprises a fully saturated (no double or triple bonds)hydrocarbon group. The alkyl group may have 1 to 25 carbon atoms(whenever it appears herein, a numerical range such as “1 to 25” refersto each integer in the given range; e.g., “1 to 25 carbon atoms” meansthat the alkyl group may consist of 1 carbon atom, 2 carbon atoms, 3carbon atoms, etc., up to and including 25 carbon atoms, although thepresent definition also covers the occurrence of the term “alkyl” whereno numerical range is designated). The alkyl group may also be a mediumsize alkyl having 1 to 15 carbon atoms. The alkyl group could also be alower alkyl having 1 to 6 carbon atoms. The alkyl group of the compoundsmay be designated as “C₄” or “C₁-C₄ alkyl” or similar designations. Byway of example only, “C₁-C₄ alkyl” indicates that there are one to fourcarbon atoms in the alkyl chain, i.e., the alkyl chain is selected frommethyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, andt-butyl. Typical alkyl groups include, but are in no way limited to,methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertiary butyl,pentyl and hexyl. The alkyl group may be substituted or unsubstituted.

As used herein, “alkenyl” refers to an alkyl group that contains in thestraight or branched hydrocarbon chain one or more double bonds. Thealkenyl group may have 2 to 25 carbon atoms (whenever it appears herein,a numerical range such as “2 to 25” refers to each integer in the givenrange; e.g., “2 to 25 carbon atoms” means that the alkenyl group mayconsist of 2 carbon atom, 3 carbon atoms, 4 carbon atoms, etc., up toand including 25 carbon atoms, although the present definition alsocovers the occurrence of the term “alkenyl” where no numerical range isdesignated). The alkenyl group may also be a medium size alkenyl having2 to 15 carbon atoms. The alkenyl group could also be a lower alkenylhaving 1 to 6 carbon atoms. The alkenyl group of the compounds may bedesignated as “C₄” or “C₂-C₄ alkyl” or similar designations. An alkenylgroup may be unsubstituted or substituted.

As used herein, “alkynyl” refers to an alkyl group that contains in thestraight or branched hydrocarbon chain one or more triple bonds. Thealkynyl group may have 2 to 25 carbon atoms (whenever it appears herein,a numerical range such as “2 to 25” refers to each integer in the givenrange; e.g., “2 to 25 carbon atoms” means that the alkynyl group mayconsist of 2 carbon atom, 3 carbon atoms, 4 carbon atoms, etc., up toand including 25 carbon atoms, although the present definition alsocovers the occurrence of the term “alkynyl” where no numerical range isdesignated). The alkynyl group may also be a medium size alkynyl having2 to 15 carbon atoms. The alkynyl group could also be a lower alkynylhaving 2 to 6 carbon atoms. The alkynyl group of the compounds may bedesignated as “C₄” or “C₂-C₄ alkyl” or similar designations. An alkynylgroup may be unsubstituted or substituted.

As used herein, “aryl” refers to a carbocyclic (all carbon) monocyclicor multicyclic aromatic ring system (including fused ring systems wheretwo carbocyclic rings share a chemical bond) that has a fullydelocalized pi-electron system throughout all the rings. The number ofcarbon atoms in an aryl group can vary. For example, the aryl group canbe a C₆-C₁₄ aryl group, a C₆-C₁₀ aryl group, or a C₆ aryl group(although the definition of C₆-C₁₀ aryl covers the occurrence of “aryl”when no numerical range is designated). Examples of aryl groups include,but are not limited to, benzene, naphthalene and azulene. An aryl groupmay be substituted or unsubstituted.

As used herein, “aralkyl” and “aryl(alkyl)” refer to an aryl groupconnected, as a substituent, via a lower alkylene group. The aralkylgroup may have 6 to 20 carbon atoms (whenever it appears herein, anumerical range such as “6 to 20” refers to each integer in the givenrange; e.g., “6 to 20 carbon atoms” means that the aralkyl group mayconsist of 6 carbon atom, 7 carbon atoms, 8 carbon atoms, etc., up toand including 20 carbon atoms, although the present definition alsocovers the occurrence of the term “aralkyl” where no numerical range isdesignated). The lower alkylene and aryl group of an aralkyl may besubstituted or unsubstituted. Examples include but are not limited tobenzyl, 2-phenylalkyl, 3-phenylalkyl, and naphthylalkyl.

“Lower alkylene groups” refer to a C₁-C₂₅ straight-chained alkyltethering groups, such as —CH₂— tethering groups, forming bonds toconnect molecular fragments via their terminal carbon atoms. Examplesinclude but are not limited to methylene (—CH₂—), ethylene (—CH₂CH₂—),propylene (—CH₂CH₂CH₂—), and butylene (—CH₂CH₂CH₂CH₂—). A lower alkylenegroup can be substituted by replacing one or more hydrogen of the loweralkylene group with a substituent(s) listed under the definition of“substituted.”

As used herein, “cycloalkyl” refers to a completely saturated (no doubleor triple bonds) mono- or multi-cyclic hydrocarbon ring system. Whencomposed of two or more rings, the rings may be joined together in afused fashion. Cycloalkyl groups can contain 3 to 10 atoms in thering(s) or 3 to 8 atoms in the ring(s). A cycloalkyl group may beunsubstituted or substituted. Typical cycloalkyl groups include, but arein no way limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl and cyclooctyl.

As used herein, “cycloalkenyl” refers to a mono- or multi-cyclichydrocarbon ring system that contains one or more double bonds in atleast one ring; although, if there is more than one, the double bondscannot form a fully delocalized pi-electron system throughout all therings (otherwise the group would be “aryl,” as defined herein). Whencomposed of two or more rings, the rings may be connected together in afused fashion. A cycloalkenyl group may be unsubstituted or substituted.

As used herein, “cycloalkynyl” refers to a mono- or multi-cyclichydrocarbon ring system that contains one or more triple bonds in atleast one ring. If there is more than one triple bond, the triple bondscannot form a fully delocalized pi-electron system throughout all therings. When composed of two or more rings, the rings may be joinedtogether in a fused fashion. A cycloalkynyl group may be unsubstitutedor substituted.

As used herein, “alkoxy” or “alkyloxy” refers to the formula —OR whereinR is an alkyl, an alkenyl, an alkynyl, a cycloalkyl, a cycloalkenyl or acycloalkynyl as defined above. A non-limiting list of alkoxys includesmethoxy, ethoxy, n-propoxy, 1-methylethoxy (isopropoxy), n-butoxy,iso-butoxy, sec-butoxy and tert-butoxy. An alkoxy may be substituted orunsubstituted.

As used herein, “acyl” refers to a hydrogen, alkyl, alkenyl, alkynyl,aryl, or heteroaryl connected, as substituents, via a carbonyl group.Examples include formyl, acetyl, propanoyl, benzoyl, and acryl. An acylmay be substituted or unsubstituted.

As used herein, “alkoxyalkyl” or “alkyloxyalkyl” refers to an alkoxygroup connected, as a substituent, via a lower alkylene group. Examplesinclude alkyl-O-alkyl- and alkoxy-alkyl- with the terms alkyl and alkoxydefined herein.

As used herein, “hydroxyalkyl” refers to an alkyl group in which one ormore of the hydrogen atoms are replaced by a hydroxy group. Exemplaryhydroxyalkyl groups include but are not limited to, 2-hydroxyethyl,3-hydroxypropyl, 2-hydroxypropyl, and 2,2-dihydroxyethyl. A hydroxyalkylmay be substituted or unsubstituted.

As used herein, “haloalkyl” refers to an alkyl group in which one ormore of the hydrogen atoms are replaced by a halogen (e.g.,mono-haloalkyl, di-haloalkyl and tri-haloalkyl). Such groups include butare not limited to, chloromethyl, fluoromethyl, difluoromethyl,trifluoromethyl and 1-chloro-2-fluoromethyl, 2-fluoroisobutyl. Ahaloalkyl may be substituted or unsubstituted.

The term “amino” as used herein refers to a —NH₂ group.

As used herein, the term “hydroxy” refers to a —OH group.

A “cyano” group refers to a “CN” group.

A “carbonyl” or an “oxo” group refers to a C═O group.

The term “azido” as used herein refers to a —N₃ group.

As used herein, “aminoalkyl” refers to an amino group connected, as asubstituent, via a lower alkylene group. Examples include H₂N-alkyl-with the term alkyl defined herein.

As used herein, “alkylcarboxyalkyl” refers to an alkyl group connected,as a substituent, to a carboxy group that is connected, as asubstituent, to an alkyl group. Examples include alkyl-C(═O)O-alkyl- andalkyl-O—C(═O)-alkyl- with the term alkyl as defined herein.

As used herein, “C-carboxyalkyl” refers to a carboxy group connected, asa substituent, to an alkyl group. Examples include HO—(C═O)-alkyl, withthe term alkyl as defined herein.

As used herein, “alkylaminoalkyl” refers to an alkyl group connected, asa substituent, to an amino group that is connected, as a substituent, toan alkyl group. Examples include alkyl-NH-alkyl-, with the term alkyl asdefined herein.

As used herein, “dialkylaminoalkyl” or “di(alkyl)aminoalkyl” refers totwo alkyl groups connected, each as a substituent, to an amino groupthat is connected, as a substituent, to an alkyl group. Examples include

with the term alkyl as defined herein.

As used herein, “alkylaminoalkylamino” refers to an alkyl groupconnected, as a substituent, to an amino group that is connected, as asubstituent, to an alkyl group that is connected, as a substituent, toan amino group. Examples include alkyl-NH-alkyl-NH—, with the term alkylas defined herein.

As used herein, “alkylaminoalkylaminoalkylamino” refers to an alkylgroup connected, as a substituent, to an amino group that is connected,as a substituent, to an alkyl group that is connected, as a substituent,to an amino group that is connected, as a substituent, to an alkylgroup. Examples include alkyl-NH-alkyl-NH-alkyl-, with the term alkyl asdefined herein.

As used herein, “arylaminoalkyl” refers to an aryl group connected, as asubstituent, to an amino group that is connected, as a substituent, toan alkyl group. Examples include aryl-NH-alkyl-, with the terms aryl andalkyl as defined herein.

As used herein, “aminoalkyloxy” refers to an amino group connected, as asubstituent, to an alkyloxy group. Examples include H₂N-alkyl-O— andH₂N-alkoxy- with the terms alkyl and alkoxy as defined herein.

As used herein, “aminoalkyloxyalkyl” refers to an amino group connected,as a substituent, to an alkyloxy group connected, as a substituent, toan alkyl group. Examples include H₂N-alkyl-O-alkyl- andH₂N-alkoxy-alkyl- with the terms alkyl and alkoxy as defined herein.

As used herein, “aminoalkylcarboxy” refers to an amino group connected,as a substituent, to an alkyl group connected, as a substituent, to acarboxy group. Examples include H₂N-alkyl-C(═O)O— and H₂N-alkyl-O—C(═O)—with the term alkyl as defined herein.

As used herein, “aminoalkylaminocarbonyl” refers to an amino groupconnected, as a substituent, to an alkyl group connected, as asubstituent, to an amino group connected, as a substituent, to acarbonyl group. Examples include H₂N-alkyl-NH—C(═O)— with the term alkylas defined herein.

As used herein, “aminoalkylcarboxamido” refers to an amino groupconnected, as a substituent, to an alkyl group connected, as asubstituent, to a carbonyl group connected, as a substituent to an aminogroup. Examples include H₂N-alkyl-C(═O)—NH— with the term alkyl asdefined herein.

As used herein, “azidoalkyloxy” refers to an azido group connected as asubstituent, to an alkyloxy group. Examples include N₃-alkyl-O— andN₃-alkoxy- with the terms alkyl and alkoxy as defined herein.

As used herein, “cyanoalkyloxy” refers to a cyano group connected as asubstituent, to an alkyloxy group. Examples include NC-alkyl-O— andNC-alkoxy- with the terms alkyl and alkoxy as defined herein.

As used herein, “guanidinoalkyloxy” refers to a guanidinyl groupconnected, as a substituent, to an alkyloxy group. Examples include

with the terms “alkyl” and “alkoxy” as defined herein.

As used herein, “guanidinoalkylcarboxy” refers to a guanidinyl groupconnected, as a substituent, to an alkyl group connected, as asubstituent, to a carboxy group. Examples include

with the term “alkyl” as defined herein.

As used herein, “quaternary ammonium alkylcarboxy” refers to aquaternized amino group connected, as a substituent, to an alkyl groupconnected, as a substituent, to a carboxy group. Examples include

with the term “alkyl” as defined herein.

The term “halogen atom” or “halogen” as used herein, means any one ofthe radio-stable atoms of column 7 of the Periodic Table of theElements, such as, fluorine, chlorine, bromine and iodine.

Where the numbers of substituents is not specified (e.g. haloalkyl),there may be one or more substituents present. For example “haloalkyl”may include one or more of the same or different halogens.

As used herein, the term “amino acid” refers to any amino acid (bothstandard and non-standard amino acids), including, but not limited to,α-amino acids, β-amino acids, γ-amino acids and δ-amino acids. Examplesof suitable amino acids include, but are not limited to, alanine,asparagine, aspartate, cysteine, glutamate, glutamine, glycine, proline,serine, tyrosine, arginine, histidine, isoleucine, leucine, lysine,methionine, phenylalanine, threonine, tryptophan and valine. Additionalexamples of suitable amino acids include, but are not limited to,ornithine, hypusine, 2-aminoisobutyric acid, dehydroalanine,gamma-aminobutyric acid, citrulline, beta-alanine, alpha-ethyl-glycine,alpha-propyl-glycine and norleucine.

A linking group is a divalent moiety used to link one steroid to anothersteroid. In some embodiments, the linking group is used to link a firstCSA compound with a second CSA compound (which may be the same ordifferent). An example of a linking group is (C₁-C₁₀) alkyloxy-(C₁-C₁₀)alkyl.

The terms “P.G.” or “protecting group” or “protecting groups” as usedherein refer to any atom or group of atoms that is added to a moleculein order to prevent existing groups in the molecule from undergoingunwanted chemical reactions. Examples of protecting group moieties aredescribed in T. W. Greene and P. G. M. Wuts, Protective Groups inOrganic Synthesis, 3. Ed. John Wiley & Sons, 1999, and in J. F. W.McOmie, Protective Groups in Organic Chemistry Plenum Press, 1973, bothof which are hereby incorporated by reference for the limited purpose ofdisclosing suitable protecting groups. The protecting group moiety maybe chosen in such a way, that they are stable to certain reactionconditions and readily removed at a convenient stage using methodologyknown from the art. A non-limiting list of protecting groups includebenzyl; substituted benzyl; alkylcarbonyls and alkoxycarbonyls (e.g.,t-butoxycarbonyl (BOC), acetyl, or isobutyryl); arylalkylcarbonyls andarylalkoxycarbonyls (e.g., benzyloxycarbonyl); substituted methyl ether(e.g. methoxymethyl ether); substituted ethyl ether; a substitutedbenzyl ether; tetrahydropyranyl ether; silyls (e.g., trimethylsilyl,triethylsilyl, triisopropylsilyl, t-butyldimethylsilyl,tri-iso-propylsilyloxymethyl, [2-(trimethylsilyl)ethoxy]methyl ort-butyldiphenylsilyl); esters (e.g. benzoate ester); carbonates (e.g.methoxymethyl-carbonate); sulfonates (e.g. tosylate or mesylate);acyclic ketal (e.g. dimethyl acetal); cyclic ketals (e.g., 1,3-dioxane,1,3-dioxolanes, and those described herein); acyclic acetal; cyclicacetal (e.g., those described herein); acyclic hemiacetal; cyclichemiacetal; cyclic dithioketals (e.g., 1,3-dithiane or 1,3-dithiolane);orthoesters (e.g., those described herein) and triarylmethyl groups(e.g., trityl; monomethoxytrityl (MMTr); 4,4′-dimethoxytrityl (DMTr);4,4′,4″-trimethoxytrityl (TMTr); and those described herein).Amino-protecting groups are known to those skilled in the art. Ingeneral, the species of protecting group is not critical, provided thatit is stable to the conditions of any subsequent reaction(s) on otherpositions of the compound and can be removed at the appropriate pointwithout adversely affecting the remainder of the molecule. In addition,a protecting group may be substituted for another after substantivesynthetic transformations are complete. Clearly, where a CSA compounddiffers from a compound disclosed herein only in that one or moreprotecting groups of the disclosed compound has been substituted with adifferent protecting group, that compound is within the disclosure

CSA compounds may also include a tether or “tail moiety” attached to thesterol backbone. The tail moiety may have variable chain length or sizeand may be one of charged, uncharged, polar, non-polar, hydrophobic,amphipathic, and the like. In various embodiments, a tail moiety may beattached at R₁₇. A tail moiety may include the heteroatom (O or N)covalently coupled to the sterol backbone. The tail moiety may, forexample, be configured to alter the hydrophobicity/hydrophilicity of theCSA compound. CSA compounds of the present disclosure having differentdegrees of hydrophobicity/hydrophilicity may, for example, havedifferent rates of uptake into different target microbes. Likewise,altering the hydrophobicity/hydrophilicity of the CSA compoundsdescribed herein may affect the retention of the CSA compounds incertain media.

B. CSA Compounds

CSA Compounds useful in accordance with this disclosure are describedherein, both generically and with particularity, and in U.S. Pat. Nos.6,350,738, 6,486,148, 6,767,904, 7,598,234, 7,754,705, U.S. ApplicationSer. Nos. 61/786,301, 13/288,892, 61/642,431, 13/554,930, 61/572,714,13/594,608, 61/576,903, 13/594,612, 13/288,902, 61/605,639, 13/783,131,61/605,642, 13/783,007, 61/132,361, 13/000,010, 61/534,185, 13/615,244,61/534,194, 13/615,324, 61/534,205, 61/637,402, 13/841,549, 61/715,277,PCT/US13/37615, 61/749,800, 61/794,721, and 61/814,816, which areincorporated herein by reference. Additional compounds are generally andspecifically described in relation to the methods discussed herein. Theskilled artisan will recognize the compounds within the generic formulaeset forth herein and understand their preparation in view of thereferences cited herein and the Examples.

In some embodiments, the CSA compound is a compound of Formula (I) or apharmaceutically acceptable salt thereof:

wherein:

rings A, B, C, and D are independently saturated, or are fully orpartially unsaturated, provided that at least two of rings A, B, C, andD are saturated;

m, n, p, and q are independently 0 or 1;

R₁ through R₄, R₆, R₇, R₁₁, R₁₂, R₁₅, R₁₆, and R₁₈ are independentlyselected from the group consisting of hydrogen, hydroxyl, a substitutedor unsubstituted alkyl, a substituted or unsubstituted hydroxyalkyl, asubstituted or unsubstituted alkyloxyalkyl, a substituted orunsubstituted alkylcarboxyalkyl, a substituted or unsubstitutedalkylaminoalkyl, a substituted or unsubstituted alkylaminoalkylamino, asubstituted or unsubstituted alkylaminoalkylaminoalkylamino, asubstituted or unsubstituted aminoalkyl, a substituted or unsubstitutedaryl, a substituted or unsubstituted arylaminoalkyl, a substituted orunsubstituted haloalkyl, a substituted or unsubstituted alkenyl, asubstituted or unsubstituted alkynyl, oxo, a linking group attached to asecond steroid, a substituted or unsubstituted aminoalkyloxy, asubstituted or unsubstituted aminoalkyloxyalkyl, a substituted orunsubstituted aminoalkylcarboxy, a substituted or unsubstitutedaminoalkylaminocarbonyl, a substituted or unsubstitutedaminoalkylcarboxamido, a substituted or unsubstituteddi(alkyl)aminoalkyl, H₂N—HC(Q₅)—C(O)—O—, H₂N—HC(Q₅)—C(O)—N(H)—, asubstituted or unsubstituted azidoalkyloxy, a substituted orunsubstituted cyanoalkyloxy, P.G.-HN—HC(Q₅)—C(O)—O—, a substituted orunsubstituted guanidinoalkyloxy, a substituted or unsubstitutedquaternaryammoniumalkylcarboxy, and a substituted or unsubstitutedguanidinoalkyl carboxy, where Q₅ is a side chain of any amino acid(including a side chain of glycine, i.e., H), and P.G. is an aminoprotecting group; and

R₅, R₈, R₉, R₁₀, R₁₃, R₁₄ and R₁₇ are independently deleted when one ofrings A, B, C, or D is unsaturated so as to complete the valency of thecarbon atom at that site, or R₅, R₈, R₉, R₁₀, R₁₃, and R₁₄ areindependently selected from the group consisting of hydrogen, hydroxyl,a substituted or unsubstituted alkyl, a substituted or unsubstitutedhydroxyalkyl, a substituted or unsubstituted alkyloxyalkyl, asubstituted or unsubstituted aminoalkyl, a substituted or unsubstitutedaryl, a substituted or unsubstituted haloalkyl, a substituted orunsubstituted alkenyl, a substituted or unsubstituted alkynyl, oxo, alinking group attached to a second steroid, a substituted orunsubstituted aminoalkyloxy, a substituted or unsubstitutedaminoalkylcarboxy, a substituted or unsubstitutedaminoalkylaminocarbonyl, a substituted or unsubstituteddi(alkyl)aminoalkyl, H₂N—HC(Q₅)—C(O)—O—, H₂N—HC(Q₅)—C(O)—N(H)—,azidoalkyloxy, cyanoalkyloxy, P.G.-HN—HC(Q₅)-C(O)—O—, guanidinoalkyloxy,and guanidine-alkylcarboxy, where Q₅ is a side chain of any amino acid,P.G. is an amino protecting group,

provided that at least two or three of R₁₋₄, R₆, R₇, R₁₁, R₁₂, R₁₅, R₁₆,R₁₇, and R₁₈ are independently selected from the group consisting of asubstituted or unsubstituted aminoalkyl, a substituted or unsubstitutedaminoalkyloxy, a substituted or unsubstituted alkylcarboxyalkyl, asubstituted or unsubstituted alkylaminoalkylamino, a substituted orunsubstituted alkylaminoalkylaminoalkylamino, a substituted orunsubstituted aminoalkylcarboxy, a substituted or unsubstitutedarylaminoalkyl, a substituted or unsubstitutedaminoalkyloxyaminoalkylaminocarbonyl, a substituted or unsubstitutedaminoalkylaminocarbonyl, a substituted or unsubstitutedaminoalkylcarboxyamido, a quaternaryammoniumalkylcarboxy, a substitutedor unsubstituted di(alkyl)aminoalkyl, H₂N—HC(Q₅)-C(O)—O—,H₂N—HC(Q₅)-C(O)—N(H)—, azidoalkyloxy, cyanoalkyloxy,P.G.-HN—HC(Q₅)-C(O)—O—, a substituted or unsubstitutedguanidinoalkyloxy, and a substituted or unsubstitutedguanidinoalkylcarboxy.

In some embodiments, R₁ through R₄, R₆, R₇, R₁₁, R₁₂, R₁₅, R₁₆, and R₁₈are independently selected from the group consisting of hydrogen,hydroxyl, a substituted or unsubstituted (C₁-C₂₂) alkyl, a substitutedor unsubstituted (C₁-C₂₂) hydroxyalkyl, a substituted or unsubstituted(C₁-C₂₂) alkyloxy-(C₁-C₂₂) alkyl, a substituted or unsubstituted(C₁-C₂₂) alkylcarboxy-(C₁-C₂₂) alkyl, a substituted or unsubstituted(C₁-C₂₂) alkylamino-(C₁-C₂₂) alkyl, a substituted or unsubstituted(C₁-C₂₂) alkylamino-(C₁-C₂₂) alkylamino, a substituted or unsubstituted(C₁-C₂₂) alkylamino-(C₁-C₂₂) alkylamino-(C₁-C₂₂) alkylamino, asubstituted or unsubstituted (C₁-C₂₂) aminoalkyl, a substituted orunsubstituted aryl, a substituted or unsubstituted arylamino-(C₁-C₂₂)alkyl, a substituted or unsubstituted (C₁-C₂₂) haloalkyl, a substitutedor unsubstituted C₂-C₆ alkenyl, a substituted or unsubstituted C₂-C₆alkynyl, oxo, a linking group attached to a second steroid, asubstituted or unsubstituted (C₁-C₂₂) aminoalkyloxy, a substituted orunsubstituted (C₁-C₂₂) aminoalkyloxy-(C₁-C₂₂) alkyl, a substituted orunsubstituted (C₁-C₂₂) aminoalkylcarboxy, a substituted or unsubstituted(C₁-C₂₂) aminoalkylaminocarbonyl, a substituted or unsubstituted(C₁-C₂₂) aminoalkylcarboxamido, a substituted or unsubstituted di(C₁-C₂₂alkyl)aminoalkyl, H₂N—HC(Q₅)-C(O)—O—, H₂N—HC(Q₅)-C(O)—N(H)—, asubstituted or unsubstituted (C₁-C₂₂) azidoalkyloxy, a substituted orunsubstituted (C₁-C₂₂) cyanoalkyloxy, P.G.-HN—HC(Q₅)-C(O)—O—, asubstituted or unsubstituted (C₁-C₂₂) guanidinoalkyloxy, a substitutedor unsubstituted (C₁-C₂₂) quaternaryammoniumalkylcarboxy, and asubstituted or unsubstituted (C₁-C₂₂) guanidinoalkylcarboxy, where Q₅ isa side chain of any amino acid (including a side chain of glycine, i.e.,H), and P.G. is an amino protecting group; R₅, R₈, R₉, R₁₀, R₁₃, R₁₄ andR₁₇ are independently deleted when one of rings A, B, C, or D isunsaturated so as to complete the valency of the carbon atom at thatsite, or R₅, R₈, R₉, R₁₀, R₁₃, and R₁₄ are independently selected fromthe group consisting of hydrogen, hydroxyl, a substituted orunsubstituted (C₁-C₂₂) alkyl, a substituted or unsubstituted (C₁-C₂₂)hydroxyalkyl, a substituted or unsubstituted (C₁-C₂₂) alkyloxy-(C₁-C₂₂)alkyl, a substituted or unsubstituted (C₁-C₂₂) aminoalkyl, a substitutedor unsubstituted aryl, a substituted or unsubstituted (C₁-C₂₂)haloalkyl, a substituted or unsubstituted (C₂-C₆) alkenyl, a substitutedor unsubstituted (C₂-C₆) alkynyl, oxo, a linking group attached to asecond steroid, a substituted or unsubstituted (C₁-C₂₂) aminoalkyloxy, asubstituted or unsubstituted (C₁-C₂₂) aminoalkylcarboxy, a substitutedor unsubstituted (C₁-C₂₂) aminoalkylaminocarbonyl, a substituted orunsubstituted di(C₁-C₂₂ alkyl)aminoalkyl, H₂N—HC(Q₅)-C(O)—O—,H₂N—HC(Q₅)-C(O)—N(H)—, a substituted or unsubstituted (C₁-C₂₂)azidoalkyloxy, a substituted or unsubstituted (C₁-C₂₂) cyanoalkyloxy,P.G.-HN—HC(Q₅)-C(O)—O—, a substituted or unsubstituted (C₁-C₂₂)guanidinoalkyloxy, and (C₁-C₂₂) guanidinoalkylcarboxy, where Q5 is aside chain of any amino acid, and P.G. is an amino protecting group;provided that at least two or three of R₁₋₄, R₆, R₇, R₁₁, R₁₂, R₁₅, R₁₆,R₁₇, and R₁₈ are independently selected from the group consisting of asubstituted or unsubstituted (C₁-C₂₂) aminoalkyl, a substituted orunsubstituted (C₁-C₂₂) aminoalkyloxy, a substituted or unsubstituted(C₁-C₂₂) alkylcarboxy-(C₁-C₂₂) alkyl, a substituted or unsubstituted(C₁-C₂₂) alkylamino-(C₁-C₂₂) alkylamino, a substituted or unsubstituted(C₁-C₂₂) alkylamino-(C₁-C₂₂) alkylamino (C₁-C₂₂) alkylamino, asubstituted or unsubstituted (C₁-C₂₂) aminoalkylcarboxy, a substitutedor unsubstituted arylamino (C₁-C₂₂) alkyl, a substituted orunsubstituted (C₁-C₂₂)aminoalkyloxy (C₁-C₂₂) aminoalkylaminocarbonyl, asubstituted or unsubstituted (C₁-C₂₂) aminoalkylaminocarbonyl, asubstituted or unsubstituted (C₁-C₂₂) aminoalkylcarboxyamido, asubstituted or unsubstituted (C₁-C₂₂) quaternaryammoniumalkylcarboxy, asubstituted or unsubstituted di(C₁-C₂₂ alkyl)aminoalkyl,H₂N—HC(Q₅)-C(O)—O—, H₂N—HC(Q₅)-C(O)—N(H)—, a substituted orunsubstituted (C₁-C₂₂) azidoalkyloxy, a substituted or unsubstituted(C₁-C₂₂) cyanoalkyloxy, P.G.-HN—HC(Q₅)-C(O)—O—, a substituted orunsubstituted (C₁-C₂₂) guanidinoalkyloxy, and a substituted orunsubstituted (C₁-C₂₂) guanidinoalkylcarboxy.

In some embodiments, R₁ through R₄, R₆, R₇, R₁₁, R₁₂, R₁₅, R₁₆, and R₁₈are independently selected from the group consisting of hydrogen,hydroxyl, an unsubstituted (C₁-C₁₈) alkyl, unsubstituted (C₁-C₁₈)hydroxyalkyl, unsubstituted (C₁-C₁₈) alkyloxy-(C₁-C₁₈) alkyl,unsubstituted (C₁-C₁₈) alkylcarboxy-(C₁-C₁₈) alkyl, unsubstituted(C₁-C₁₈) alkylamino-(C₁-C₁₈)alkyl, unsubstituted (C₁-C₁₈)alkylamino-(C₁-C₁₈) alkylamino, (C₁-C₁₈) alkylamino-(C₁-C₁₈)alkylamino-(C₁-C₁₈) alkylamino, an unsubstituted (C₁-C₁₈) aminoalkyl, anunsubstituted aryl, an unsubstituted arylamino-(C₁-C₁₈) alkyl, oxo, anunsubstituted (C₁-C₁₈) aminoalkyloxy, an unsubstituted (C₁-C₁₈)aminoalkyloxy-(C₁-C₁₈) alkyl, an unsubstituted (C₁-C₁₈)aminoalkylcarboxy, an unsubstituted (C₁-C₁₈) aminoalkylaminocarbonyl, anunsubstituted (C₁-C₁₈) aminoalkylcarboxamido, an unsubstituted di(C₁-C₁₈alkyl)aminoalkyl, unsubstituted (C₁-C₁₈) guanidinoalkyloxy,unsubstituted (C₁-C₁₈) quaternaryammoniumalkylcarboxy, and unsubstituted(C₁-C₁₈) guanidinoalkylcarboxy; R₅, R₈, R₉, R₁₀, R₁₃, R₁₄ and R₁₇ areindependently deleted when one of rings A, B, C, or D is unsaturated soas to complete the valency of the carbon atom at that site, or R₅, R₈,R₉, R₁₀, R₁₃, and R₁₄ are independently selected from the groupconsisting of hydrogen, hydroxyl, an unsubstituted (C₁-C₁₈) alkyl,unsubstituted (C₁-C₁₈) hydroxyalkyl, unsubstituted (C₁-C₁₈)alkyloxy-(C₁-C₁₈) alkyl, unsubstituted (C₁-C₁₈) alkylcarboxy-(C₁-C₁₈)alkyl, unsubstituted (C₁-C₁₈) alkylamino-(C₁-C₁₈)alkyl, (C₁-C₁₈)alkylamino-(C₁-C₁₈) alkylamino, unsubstituted (C₁-C₁₈)alkylamino-(C₁-C₁₈) alkylamino-(C₁-C₁₈) alkylamino, an unsubstituted(C₁-C₁₈) aminoalkyl, an unsubstituted aryl, an unsubstitutedarylamino-(C₁-C₁₈) alkyl, oxo, an unsubstituted (C₁-C₁₈) aminoalkyloxy,an unsubstituted (C₁-C₁₈) aminoalkyloxy-(C₁-C₁₈) alkyl, an unsubstituted(C₁-C₁₈) aminoalkylcarboxy, an unsubstituted (C₁-C₁₈)aminoalkylaminocarbonyl, an unsubstituted (C₁-C₁₈)aminoalkylcarboxamido, an unsubstituted di(C₁-C₁₈ alkyl)aminoalkyl,unsubstituted (C₁-C₁₈) guanidinoalkyloxy, unsubstituted (C₁-C₁₈)quaternaryammoniumalkylcarboxy, and unsubstituted (C₁-C₁₈)guanidinoalkyl carboxy; provided that at least two or three of R₁₋₄, R₆,R₇, R₁₁, R₁₂, R₁₅, R₁₆, R₁₇, and R₁₈ are independently selected from thegroup consisting of of hydrogen, hydroxyl, an unsubstituted (C₁-C₁₈)alkyl, unsubstituted (C₁-C₁₈) hydroxyalkyl, unsubstituted (C₁-C₁₈)alkyloxy-(C₁-C₁₈) alkyl, unsubstituted (C₁-C₁₈) alkylcarboxy-(C₁-C₁₈)alkyl, unsubstituted (C₁-C₁₈) alkylamino-(C₁-C₁₈)alkyl, unsubstituted(C₁-C₁₈) alkylamino-(C₁-C₁₈) alkylamino, unsubstituted (C₁-C₁₈)alkylamino-(C₁-C₁₈) alkylamino-(C₁-C₁₈) alkylamino, an unsubstituted(C₁-C₁₈) aminoalkyl, an unsubstituted aryl, an unsubstitutedarylamino-(C₁-C₁₈) alkyl, oxo, an unsubstituted (C₁-C₁₈) aminoalkyloxy,an unsubstituted (C₁-C₁₈) aminoalkyloxy-(C₁-C₁₈) alkyl, an unsubstituted(C₁-C₁₈) aminoalkylcarboxy, an unsubstituted (C₁-C₁₈)aminoalkylaminocarbonyl, an unsubstituted (C₁-C₁₈)aminoalkylcarboxamido, an unsubstituted di(C₁-C₁₈ alkyl) amino alkyl,unsubstituted (C₁-C₁₈) guanidinoalkyloxy, unsubstituted (C₁-C₁₈)quaternaryammoniumalkylcarboxy, and unsubstituted (C₁-C₁₈)guanidinoalkyl carboxy.

In some embodiments, the CSA compound, or a pharmaceutically acceptablesalt thereof, is selected from the compound of Formula (IA), which is asubgenus of Formula (I) in that R₁₅ is omitted:

In some embodiments, rings A, B, C, and D are independently saturated.

In some embodiments, one or more of rings A, B, C, and D areheterocyclic.

In some embodiments, rings A, B, C, and D are non-heterocyclic.

In some embodiments, R₃, R₇, R₁₂, and R₁₈ are independently selectedfrom the group consisting of hydrogen, an unsubstituted (C₁-C₁₈) alkyl,unsubstituted (C₁-C₁₈) hydroxyalkyl, unsubstituted (C₁-C₁₈)alkyloxy-(C₁-C₁₈) alkyl, unsubstituted (C₁-C₁₈) alkylcarboxy-(C₁-C₁₈)alkyl, unsubstituted (C₁-C₁₈) alkylamino-(C₁-C₁₈)alkyl, unsubstituted(C₁-C₁₈) alkylamino-(C₁-C₁₈) alkylamino, unsubstituted (C₁-C₁₈)alkylamino-(C₁-C₁₈) alkylamino-(C₁-C₁₈) alkylamino, an unsubstituted(C₁-C₁₈) aminoalkyl, an unsubstituted arylamino-(C₁-C₁₈) alkyl, anunsubstituted (C₁-C₁₈) aminoalkyloxy, an unsubstituted (C₁-C₁₈)aminoalkyloxy-(C₁-C₁₈) alkyl, an unsubstituted (C₁-C₁₈)aminoalkylcarboxy, an unsubstituted (C₁-C₁₈) amino alkyl-aminocarbonyl,an unsubstituted (C₁-C₁₈) aminoalkylcarboxamido, an unsubstituteddi(C₁-C₁₈ alkyl)aminoalkyl, unsubstituted (C₁-C₁₈) guanidinoalkyloxy,unsubstituted (C₁-C₁₈) quaternaryammoniumalkylcarboxy, and unsubstituted(C₁-C₁₈) guanidinoalkyl carboxy; and R₁, R₂, R₄, R₅, R₆, R₈, R₉, R₁₀,R₁₁, R₁₃, R₁₄, R₁₅, R₁₆, and R₁₇ are independently selected from thegroup consisting of hydrogen and unsubstituted (C₁-C₆) alkyl.

In some embodiments, R₃, R₇, R₁₂, and R₁₈ are independently selectedfrom the group consisting of hydrogen, an unsubstituted (C₁-C₆) alkyl,unsubstituted (C₁-C₆) hydroxyalkyl, unsubstituted (C₁-C₁₆)alkyloxy-(C₁-C₅) alkyl, unsubstituted (C₁-C₁₆) alkylcarboxy-(C₁-C₅)alkyl, unsubstituted (C₁-C₁₆) alkylamino-(C₁-C₅)alkyl, unsubstituted(C₁-C₁₆) alkylamino-(C₁-C₅) alkylamino, unsubstituted (C₁-C₁₆)alkylamino-(C₁-C₁₆) alkylamino-(C₁-C₅) alkylamino, an unsubstituted(C₁-C₁₆) aminoalkyl, an unsubstituted arylamino-(C₁-C₅) alkyl, anunsubstituted (C₁-C₅) aminoalkyloxy, an unsubstituted (C₁-C₁₆)aminoalkyloxy-(C₁-C₅) alkyl, an unsubstituted (C₁-C₅) aminoalkylcarboxy,an unsubstituted (C₁-C₅) aminoalkylaminocarbonyl, an unsubstituted(C₁-C₅) aminoalkylcarboxamido, an unsubstituted di(C₁-C₅alkyl)amino-(C₁-C₅) alkyl, unsubstituted (C₁-C₅) guanidinoalkyloxy,unsubstituted (C₁-C₁₆) quaternaryammoniumalkylcarboxy, and unsubstituted(C₁-C₁₆) guanidinoalkylcarboxy.

In some embodiments, R₁, R₂, R₄, R₅, R₆, R₈, R₁₀, R₁₁, R₁₄, R₁₆, and R₁₇are each hydrogen; and R₉ and R₁₃ are each methyl.

In some embodiments, R₃, R₇, R₁₂, and R₁₈ are independently selectedfrom the group consisting of aminoalkyloxy; aminoalkylcarboxy;alkylaminoalkyl; alkoxycarbonylalkyl; alkylcarbonylalkyl;di(alkyl)aminoalkyl; alkylcarboxyalkyl; and hydroxyalkyl.

In some embodiments, R₃, R₇, and R₁₂ are independently selected from thegroup consisting of aminoalkyloxy and aminoalkylcarboxy; and R₁₈ isselected from the group consisting of alkylaminoalkyl;alkoxycarbonylalkyl; alkylcarbonyloxyalkyl; di(alkyl)aminoalkyl;alkylaminoalkyl; alkyoxycarbonylalkyl; alkylcarboxyalkyl; andhydroxyalkyl.

In some embodiments, R₃, R₇, and R₁₂ are the same. In some embodiments,R₃, R₇, and R₁₂ are aminoalkyloxy. In some embodiments, R₃, R₇, and R₁₂are aminoalkylcarboxy. In some embodiments, R₁₈ is alkylaminoalkyl. Insome embodiments, R₁₈ is alkoxycarbonylalkyl. In some embodiments, R₁₈is di(alkyl)aminoalkyl. In some embodiments, R₁₈ is alkylcarboxyalkyl.In some embodiments, R₁₈ is hydroxyalkyl.

In some embodiments, R₃, R₇, R₁₂, and R₁₈ are independently selectedfrom the group consisting of amino-C₃-alkyloxy; amino-C₃-alkyl-carboxy;C₈-alkylamino-C₅-alkyl; C₈-alkoxy-carbonyl-C₄-alkyl;C₈-alkyl-carbonyl-C₄-alkyl; di-(C₅-alkyl)amino-C₅-alkyl;C₁₃-alkylamino-C₅-alkyl; C₆-alkoxy-carbonyl-C₄-alkyl;C₆-alkyl-carboxy-C₄-alkyl; and C₁₆-alkylamino-C₅-alkyl.

In some embodiments, at least two, or at least three, of m, n, p, and qare 1. In some embodiments, m, n, and p are each 1 and q is 0.

In some embodiments, the CSA compound, or a pharmaceutically acceptablesalt thereof, is selected from the compound of Formula (IB), which is asubgenus of Formula (IA):

In some embodiments, the CSA compound, or a pharmaceutically acceptablesalt thereof of the compound of Formula (IB), is selected from the groupconsisting of:

In some embodiments, the CSA compound, or a pharmaceutically acceptablesalt thereof, is selected from the compound of Formula (II), which isrelated to, but not identical to, Formula (I), e.g., in that R₁₈, ratherthan R₁₅, is optional and can be omitted:

wherein

rings A, B, C, and D are independently saturated, or are fully orpartially unsaturated, provided that at least two of rings A, B, C, andD are saturated;

m, n, p, and q are independently 0 or 1;

each of R₁ through R₄, R₆, R₇, R₁₁, R₁₂, R₁₅, R₁₆, R₁₇, and R₁₈ isindependently selected from the group consisting of hydrogen, hydroxyl,a substituted or unsubstituted (C₁-C₁₀) alkyl, (C₁-C₁₀) hydroxyalkyl,(C₁-C₁₀) alkyloxy-(C₁-C₁₀) alkyl, (C₁-C₁₀) alkylcarboxy-(C₁-C₁₀) alkyl,(C₁-C₁₀) alkylamino-(C₁-C₁₀) alkyl, (C₁-C₁₀) alkylamino-(C₁-C₁₀)alkylamino, (C₁-C₁₀) alkylamino-(C₁-C₁₀) alkylamino-(C₁-C₁₀) alkylamino,a substituted or unsubstituted (C₁-C₁₀) aminoalkyl, a substituted orunsubstituted aryl, a substituted or unsubstituted arylamino-(C₁-C₁₀)alkyl, (C₁-C₁₀) haloalkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, oxo, a linkinggroup attached to a second steroid, a substituted or unsubstituted(C₁-C₁₀) aminoalkyloxy, a substituted or unsubstituted (C₁-C₁₀)aminoalkyloxy-(C₁-C₁₀) alkyl, a substituted or unsubstituted (C₁-C₁₀)aminoalkylcarboxy, a substituted or unsubstituted (C₁-C₁₀)aminoalkylaminocarbonyl, a substituted or unsubstituted(C₁-C₁₀)aminoalkylcarboxamido, H₂N—HC(Q₅)-C(O)—O, H₂N—HC(Q₅)-C(O)—N(H)—,(C₁-C₁₀) azidoalkyloxy, (C₁-C₁₀) cyanoalkyloxy, P.G.-HN—HC(Q₅)-C(O)—O—,(C₁-C₁₀) guanidinoalkyloxy, (C₁-C₁₀) quaternary ammonium alkylcarboxy,and (C₁-C₁₀) guanidinoalkyl carboxy, where Q₅ is a side chain of anyamino acid (including a side chain of glycine, i.e., H), P.G. is anamino protecting group; and

each of R₅, R₈, R₉, R₁₀, R₁₃, and R₁₄ may be independently deleted whenone of fused rings A, B, C, or D is unsaturated so as to complete thevalency of the carbon atom at that site, or selected from the groupconsisting of hydrogen, hydroxyl, a substituted or unsubstituted(C₁-C₁₀) alkyl, (C₁-C₁₀) hydroxyalkyl, (C₁-C₁₀) alkyloxy-(C₁-C_(m))alkyl, a substituted or unsubstituted (C₁-C₁₀) aminoalkyl, a substitutedor unsubstituted aryl, (C₁-C₁₀) haloalkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,oxo, a linking group attached to a second steroid, a substituted orunsubstituted (C₁-C₁₀) aminoalkyloxy, a substituted or unsubstituted(C₁-C₁₀) aminoalkylcarboxy, a substituted or unsubstituted (C₁-C₁₀)aminoalkylaminocarbonyl, H₂N—HC(Q₅)-C(O)—O—, H₂N—HC(Q₅)-C(O)—N(H)—,(C₁-C₁₀)azidoalkyloxy, (C₁-C₁₀) cyanoalkyloxy, P.G.-HN—HC(Q₅)-C(O)—O—,(C₁-C₁₀) guanidinoalkyloxy, and (C₁-C₁₀) guanidinoalkyl carboxy, whereQ₅ is a side chain of any amino acid, P.G. is an amino protecting group,provided that at least two or three of R₁₋₄, R₆, R₇, R₁₁, R₁₂, R₁₅, R₁₆,R₁₇, and R₁₈ are independently selected from the group consisting of asubstituted or unsubstituted (C₁-C₁₀) aminoalkyl, a substituted orunsubstituted (C₁-C₁₀) aminoalkyloxy, (C₁-C₁₀) alkylcarboxy-(C₁-C₁₀)alkyl, (C₁-C₁₀) alkylamino-(C₁-C₁₀) alkylamino, (C₁-C₁₀)alkylamino-(C₁-C₁₀) alkylamino-(C₁-C₁₀) alkylamino, a substituted orunsubstituted (C₁-C₁₀) aminoalkylcarboxy, a substituted or unsubstitutedarylamino (C₁-C₁₀) alkyl, a substituted or unsubstituted(C₁-C₁₀)aminoalkyloxy-(C₁-C₁₀) aminoalkylaminocarbonyl, a substituted orunsubstituted (C₁-C₁₀) aminoalkylaminocarbonyl, a substituted orunsubstituted (C₁-C₅) aminoalkylcarboxyamido, a (C₁-C₁₀) quaternaryammonium alkylcarboxy, H₂N—HC(Q₅)-C(O)—O—, H₂N—HC(Q₅)-C(O)—N(H)—,(C₁-C₁₀) azidoalkyloxy, (C₁-C₁₀) cyanoalkyloxy, P.G.-HN—HC(Q₅)-C(O)—O—,(C₁-C₁₀) guanidine-alkyloxy, and a (C₁-C₁₀) guanidinoalkylcarboxy.

In Formula (II), at least two or three of R₃, R₇, or R₁₂ mayindependently include a cationic moiety attached to the Formula (II)structure via a hydrolysable linkage. Optionally, a tail moiety may beattached to Formula (II) at R₁₇. The tail moiety may be charged,uncharged, polar, non-polar, hydrophobic, amphipathic, and the like.Although not required, at least two or three of m, n, p. and q can be 1.In a preferred embodiment, m, n, and p=1 and q=0.

In some embodiments, the compound of Formula (II) or pharmaceuticallyacceptable salt can be represented by Formula (IIA), which is a subgenusof Formula (II) in that R₁₈ is omitted:

wherein

fused rings A, B, C, and D are independently saturated or fully orpartially unsaturated;

each of R₁ through R₄, R₆, R₇, R₁₁, R₁₂, R₁₅, R₁₆, and R₁₇ isindependently selected from the group consisting of hydrogen, hydroxyl,a substituted or unsubstituted (C₁-C₁₀) alkyl, (C₁-C₁₀) hydroxyalkyl,(C₁-C₁₀) alkyloxy-(C₁-C₁₀) alkyl, (C₁-C₁₀) alkylcarboxy-(C₁-C₁₀) alkyl,C₁-C₁₀) alkylamino-(C₁-C₁₀) alkyl, (C₁-C₁₀) alkylamino-(C₁-C₁₀)alkylamino, (C₁-C₁₀) alkylamino-(C₁-C₁₀) alkylamino-(C₁-C₁₀) alkylamino,a substituted or unsubstituted (C₁-C₁₀) aminoalkyl, a substituted orunsubstituted aryl, a substituted or unsubstituted arylamino-(C₁-C₁₀)alkyl, (C₁-C₁₀)haloalkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, oxo, a linkinggroup attached to a second steroid, a substituted or unsubstituted(C₁-C₁₀) aminoalkyloxy, a substituted or unsubstituted (C₁-C₁₀)aminoalkyloxy-(C₁-C₁₀) alkyl, a substituted or unsubstituted (C₁-C₁₀)aminoalkylcarboxy, a substituted or unsubstituted (C₁-C₁₀)aminoalkylaminocarbonyl, a substituted or unsubstituted (C₁-C₁₀)aminoalkylcarboxamido, H₂N—HC(Q₅)-C(O)—O—, H₂N—HC(Q₅)-C(O)—N(H)—,(C₁-C₁₀) azidoalkyloxy, (C₁-C₁₀) cyanoalkyloxy, P.G.-HN—HC(Q₅)-C(O)—O—,(C₁-C₁₀) guanidinoalkyloxy, (C₁-C₁₀) quaternary ammonium alkylcarboxy,and (C₁-C₁₀) guanidinoalkyl carboxy, where Q₅ is a side chain of anyamino acid (including the side chain of glycine, i.e., H), PG. is anamino protecting group;

R₅, R₈, R₉, R₁₀, R₁₃, and R₁₄ is each independently: deleted when one offused rings A, B, C, or D is unsaturated so as to complete the valencyof the carbon atom at that site, or selected from the group consistingof hydrogen, hydroxyl, a substituted or unsubstituted (C₁-C₁₀) alkyl,(C₁-C₁₀) hydroxyalkyl, (C₁-C₁₀) alkyloxy-(C₁-C₁₀) alkyl, a substitutedor unsubstituted (C₁-C₁₀) aminoalkyl, a substituted or unsubstitutedaryl, C₁-C₁₀ haloalkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, oxo, a linkinggroup attached to a second steroid, a substituted or unsubstituted(C₁-C₁₀) aminoalkyloxy, a substituted or unsubstituted (C₁-C₁₀)aminoalkylcarboxy, a substituted or unsubstituted (C₁-C₁₀)aminoalkylaminocarbonyl, H₂N—HC(Q₅)-C(O)—O—, H₂N—HC(Q₅)-C(O)—N(H)—,(C₁-C₁₀) azidoalkyloxy, (C₁-C₁₀) cyanoalkyloxy, P.G.-HN—HC(Q₅)-C(O)—O—,(C₁-C₁₀) guanidine-alkyloxy, and (C₁-C₁₀) guanidinoalkylcarboxy, whereQ₅ is a side chain of any amino acid, PG. is an amino protecting group;and

at least two of R₁ through R₁₄ are independently selected from the groupconsisting of a substituted or unsubstituted (C₁-C₁₀) aminoalkyloxy,(C₁-C₁₀) alkylcarboxy(C₁-C₁₀) alkyl, (C₁-C₁₀) alkylamino-(C₁-C₁₀)alkylamino, (C₁-C₁₀) alkylamino-(C₁-C₁₀) alkylamino-(C₁-C₁₀) alkylamino,a substituted or unsubstituted (C₁-C₁₀) aminoalkylcarboxy, a substitutedor unsubstituted arylamino(C₁-C₁₀) alkyl, a substituted or unsubstituted(C₁-C₁₀) amino alkyloxy-(C₁-C₁₀) alkyl, a substituted or unsubstituted(C₁-C₁₀) aminoalkylaminocarbonyl, (C₁-C₁₀) quaternary ammoniumalkylcarboxy, H₂N—HC(Q₅)—C(O)—O—, H₂N—HC(Q₅)-C(O)—N(H)—, (C₁-C₁₀)azidoalkyloxy, (C₁-C₁₀) cyanoalkyloxy, PG.-HN—HC(Q₅)-C(O)—O—, (C₁-C₁₀)guanidinoalkyloxy, and (C₁-C₁₀) guanidinoalkylcarboxy.

In some embodiments, compounds comprise a ring system of at least 4fused rings, where each of the rings has from 5-7 atoms. The ring systemhas two faces, and contains 3 chains attached to the same face. Each ofthe chains contains a nitrogen-containing group that is separated fromthe ring system by at least one atom; the nitrogen-containing group isan amino group, e.g., a primary amino group, or a guanidino group.

C. Pharmaceutically Acceptable Salts

The compounds and compositions disclosed herein are optionally preparedas pharmaceutically acceptable salts. The term “pharmaceuticallyacceptable salt” as used herein is a broad term, and is to be given itsordinary and customary meaning to a skilled artisan (and is not to belimited to a special or customized meaning), and refers withoutlimitation to a salt of a compound that does not cause significantirritation to an organism to which it is administered and does notabrogate the biological activity and properties of the compound. In someembodiments, the salt is an acid addition salt of the compound.Pharmaceutical salts can be obtained by reacting a compound withinorganic acids such as hydrohalic acid (e.g., hydrochloric acid orhydrobromic acid), sulfuric acid, nitric acid, and phosphoric acid.Pharmaceutical salts can also be obtained by reacting a compound with anorganic acid such as aliphatic or aromatic carboxylic or sulfonic acids,for example formic acid, acetic acid, propionic acid, glycolic acid,pyruvic acid, malonic acid, maleic acid, fumaric acid, trifluoroaceticacid, benzoic acid, cinnamic acid, mandelic acid, succinic acid, lacticacid, malic acid, tartaric acid, citric acid, ascorbic acid, nicotinicacid, methanesulfonic acid, ethanesulfonic acid, p-toluensulfonic acid,salicylic acid, stearic acid, muconic acid, butyric acid, phenylaceticacid, phenylbutyric acid, valproic acid, 1,2-ethanedisulfonic acid,2-hydroxyethanesulfonic acid, benzenesulfonic acid,2-naphthalenesulfonic acid, or naphthalenesulfonic acid. Pharmaceuticalsalts can also be obtained by reacting a compound with a base to form asalt such as an ammonium salt, an alkali metal salt, such as a lithium,sodium or a potassium salt, an alkaline earth metal salt, such as acalcium, magnesium or aluminum salt, a salt of organic bases such asdicyclohexylamine, N-methyl-D-glucamine, tris(hydroxymethyl)methylamine,C₁-C₇ alkylamine, cyclohexylamine, dicyclohexylamine, triethanolamine,ethylenediamine, ethanolamine, diethanolamine, triethanolamine,tromethamine, and salts with amino acids such as arginine and lysine; ora salt of an inorganic base, such as aluminum hydroxide, calciumhydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, orthe like.

In some embodiments, the pharmaceutically acceptable salt is ahydrochloride salt. In some embodiments, the pharmaceutically acceptablesalt is a mono-hydrochloride salt, a di-hydrochloride salt, atri-hydrochloride salt, or a tetra-hydrochloride salt. Additionalexamples of pharmaceutically acceptable salts include sulfuric acidaddition salts and sulfonic acid addition salts.1,5-naphthalenedisulfonic acid is a particularly useful sulfonic acidaddition salt.

D. Pharmaceutical Compositions

While it is possible for the compounds described herein to beadministered alone, it may be desirable to formulate compounds aspharmaceutical compositions. As such, in yet another aspect,pharmaceutical compositions useful in the methods and uses of thedisclosed embodiments are provided. More particularly, thepharmaceutical compositions described herein may be useful, inter alia,for treating or preventing infertility in a mammal. A pharmaceuticalcomposition is any composition that may be administered in vitro or invivo or both to a subject in order to treat or ameliorate a condition.In a preferred embodiment, a pharmaceutical composition may beadministered in vivo. A subject may include one or more cells ortissues, or organisms. In some exemplary embodiments, the subject is ananimal. In some embodiments, the animal is a mammal. A mammal includesany mammal, such as by way of non-limiting example, cattle, pigs, sheep,goats, horses, camels, buffalo, cats, dogs, rats, mice, humans, andprimates.

As used herein the terms “pharmaceutically acceptable” and“physiologically acceptable” mean a biologically compatible formulation,gaseous, liquid or solid, or mixture thereof, which is suitable for oneor more routes of administration, in vivo delivery, or contact. Aformulation is compatible in that it does not destroy activity of anactive ingredient therein (e.g., a CSA compound), or induce adverse sideeffects that far outweigh any prophylactic or therapeutic effect orbenefit.

In an embodiment, the pharmaceutical compositions may be formulated withpharmaceutically acceptable excipients such as carriers, solvents,stabilizers, adjuvants, diluents, etc., depending upon the particularmode of administration and dosage form. The pharmaceutical compositionsshould generally be formulated to achieve a physiologically compatiblepH, and may range from a pH of about 3 to a pH of about 11, preferablyabout pH 3 to about pH 7, depending on the formulation and route ofadministration. In alternative embodiments, it may be preferred that thepH is adjusted to a range from about pH 5.0 to about pH 8. Moreparticularly, the pharmaceutical compositions may comprise atherapeutically or prophylactically effective amount of at least onecompound as described herein, together with one or more pharmaceuticallyacceptable excipients. Optionally, the pharmaceutical compositions maycomprise a combination of the compounds described herein, or may includea second active ingredient useful in the treatment or prevention ofbacterial infection (e.g., antibacterial or anti-microbial agents).

Formulations, e.g., for parenteral or oral administration, are mosttypically solids, liquid solutions, emulsions or suspensions, whileinhalable formulations for pulmonary administration are generallyliquids or powders. A preferred pharmaceutical composition may also beformulated as a lyophilized solid that is reconstituted with aphysiologically compatible solvent prior to administration. Alternativepharmaceutical compositions may be formulated as syrups, creams,ointments, tablets, and the like. Lavages and/or infusions of the CSAcompositions may preferably be in the form of a liquid that isintroduced into the uterus of the female mammal.

The term “pharmaceutically acceptable excipient” refers to an excipientfor administration of a pharmaceutical agent, such as the compoundsdescribed herein. The term refers to any pharmaceutical excipient thatmay be administered without undue toxicity.

Pharmaceutically acceptable excipients are determined in part by theparticular composition being administered, as well as by the particularmethod used to administer the composition. Accordingly, there exists awide variety of suitable formulations of pharmaceutical compositions(see, e.g., Remington's Pharmaceutical Sciences).

Suitable excipients may be carrier molecules that include large, slowlymetabolized macromolecules such as proteins, polysaccharides, polylacticacids, polyglycolic acids, polymeric amino acids, amino acid copolymers,and inactive virus particles. Other exemplary excipients includeantioxidants such as ascorbic acid; chelating agents such as EDTA;carbohydrates such as dextrin, hydroxyalkylcellulose,hydroxyalkylmethylcellulose, stearic acid; liquids such as oils, water,saline, glycerol and ethanol; wetting or emulsifying agents; pHbuffering substances; and the like. Liposomes are also included withinthe definition of pharmaceutically acceptable excipients.

The pharmaceutical compositions described herein may be formulated inany form suitable for the intended method of administration. Whenintended for oral use for example, tablets, troches, lozenges, aqueousor oil suspensions, non-aqueous solutions, dispersible powders orgranules (including micronized particles or nanoparticles), emulsions,hard or soft capsules, syrups or elixirs may be prepared. Compositionsintended for oral use may be prepared according to any method known tothe art for the manufacture of pharmaceutical compositions, and suchcompositions may contain one or more agents including sweetening agents,flavoring agents, coloring agents and preserving agents, in order toprovide a palatable preparation. Pharmaceutical compositions may beformulated as suspensions comprising a compound of the embodiments inadmixture with at least one pharmaceutically acceptable excipientsuitable for the manufacture of a suspension. In yet another embodiment,pharmaceutical compositions may be formulated as dispersible powders andgranules suitable for preparation of a suspension by the addition ofsuitable excipients.

Excipients suitable for use in connection with suspensions includesuspending agents, such as sodium carboxymethylcellulose,methylcellulose, hydroxypropyl methylcellulose, sodium alginate,polyvinylpyrrolidone, gum tragacanth, gum acacia, dispersing or wettingagents such as a naturally occurring phosphatide (e.g., lecithin), acondensation product of an alkylene oxide with a fatty acid (e.g.,polyoxyethylene stearate), a condensation product of ethylene oxide witha long chain aliphatic alcohol (e.g., heptadecaethyleneoxycethanol), acondensation product of ethylene oxide with a partial ester derived froma fatty acid and a hexitol anhydride (e.g., polyoxyethylene sorbitanmonooleate); polysaccharides and polysaccharide-like compounds (e.g.dextran sulfate); glycoaminoglycans and glycosaminoglycan-like compounds(e.g., hyaluronic acid); and thickening agents, such as carbomer,beeswax, hard paraffin or cetyl alcohol. The suspensions may alsocontain one or more preservatives such as acetic acid, methyl and/orn-propyl p-hydroxy-benzoate; one or more coloring agents; one or moreflavoring agents; and one or more sweetening agents such as sucrose orsaccharin.

The pharmaceutical compositions may also be in the form of oil-in wateremulsions. The oily phase may be a vegetable oil, such as olive oil orarachis oil, a mineral oil, such as liquid paraffin, or a mixture ofthese. Suitable emulsifying agents include naturally-occurring gums,such as gum acacia and gum tragacanth; naturally occurring phosphatides,such as soybean lecithin, esters or partial esters derived from fattyacids; hexitol anhydrides, such as sorbitan monooleate; and condensationproducts of these partial esters with ethylene oxide, such aspolyoxyethylene sorbitan monooleate. The emulsion may also containsweetening and flavoring agents. Syrups and elixirs may be formulatedwith sweetening agents, such as glycerol, sorbitol or sucrose. Suchformulations may also contain a demulcent, a preservative, a flavoringor a coloring agent.

Additionally, the pharmaceutical compositions may be in the form of asterile injectable preparation, such as a sterile injectable aqueousemulsion or oleaginous suspension. This emulsion or suspension may beformulated according to the known art using those suitable dispersing orwetting agents and suspending agents which have been mentioned above.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally acceptable diluent orsolvent, such as a solution in 1,2-propanediol (propylene glycol).

The sterile injectable preparation may also be prepared as a lyophilizedpowder. Among the acceptable vehicles and solvents that may be employedare water, Ringer's solution, and isotonic sodium chloride solution. Inaddition, sterile fixed oils may be employed as a solvent or suspendingmedium. For this purpose any bland fixed oil may be employed includingsynthetic mono- or diglycerides. In addition, fatty acids such as oleicacid may likewise be used in the preparation of injectables.

To obtain a stable water-soluble dose form of a pharmaceuticalcomposition, a pharmaceutically acceptable salt of a compound describedherein may be dissolved in an aqueous solution of an organic orinorganic acid, such as 0.3 M solution of succinic acid, or morepreferably, citric acid. If a soluble salt form is not available, thecompound may be dissolved in a suitable co-solvent or combination ofco-solvents. Examples of suitable co-solvents include alcohol, propyleneglycol, polyethylene glycol 300, polysorbate 80, glycerin and the likein concentrations ranging from about 0 to about 60% of the total volume.In applications where DMSO is not prohibited, the active may first bedissolved in DMSO and then diluted with water.

The pharmaceutical composition may also be in the form of a solution ofa salt form of the active ingredient in an appropriate aqueous vehicle,such as water or isotonic saline or dextrose solution. Also contemplatedare compounds which have been modified by substitutions or additions ofchemical or biochemical moieties which make them more suitable fordelivery (e.g., increase solubility, bioactivity, palatability, decreaseadverse reactions, etc.), for example by esterification, glycosylation,PEGylation, etc.

In one embodiment, the compounds described herein may be formulated fororal administration in a lipid-based formulation suitable for lowsolubility compounds. Lipid-based formulations can generally enhance theoral bioavailability of such compounds.

As such, a pharmaceutical composition comprises a therapeutically orprophylactically effective amount of a compound described herein,together with at least one pharmaceutically acceptable excipientselected from the group consisting of—medium chain fatty acids orpropylene glycol esters thereof (e.g., propylene glycol esters of ediblefatty acids such as caprylic and capric fatty acids) andpharmaceutically acceptable surfactants such as Polyoxyl 40 hydrogenatedcastor oil.

In an alternative preferred embodiment, cyclodextrins may be added asaqueous solubility enhancers. Preferred cyclodextrins includehydroxypropyl, hydroxyethyl, glucosyl, maltosyl and maltotriosylderivatives of α-, β-, and γ-cyclodextrin. A particularly preferredcyclodextrin solubility enhancer is hydroxypropyl-o-cyclodextrin (BPBC),which may be added to any of the above-described compositions to furtherimprove the aqueous solubility characteristics of the compounds of theembodiments. In one embodiment, the composition comprises about 0.1% toabout 20% hydroxypropyl-o-cyclodextrin, more preferably about 1% toabout 15% hydroxypropyl-o-cyclodextrin, and even more preferably fromabout 2.5% to about 10% hydroxypropyl-o-cyclodextrin. The amount ofsolubility enhancer employed will depend on the amount of the compoundof the embodiments in the composition.

In some exemplary embodiments, a CSA comprises a multimer (e.g., adimer, trimer, tetramer, or higher order polymer). In some exemplaryembodiments, the CSAs can be incorporated into pharmaceuticalcompositions or formulations. Such pharmaceuticalcompositions/formulations are useful for administration to a subject, invivo or ex vivo. Pharmaceutical compositions and formulations includecarriers or excipients for administration to a subject.

Such formulations include solvents (aqueous or non-aqueous), solutions(aqueous or non-aqueous), emulsions (e.g., oil-in-water orwater-in-oil), suspensions, syrups, elixirs, dispersion and suspensionmedia, coatings, isotonic and absorption promoting or delaying agents,compatible with pharmaceutical administration or in vivo contact ordelivery. Aqueous and non-aqueous solvents, solutions and suspensionsmay include suspending agents and thickening agents. Suchpharmaceutically acceptable carriers include tablets (coated oruncoated), capsules (hard or soft), microbeads, powder, granules andcrystals. Supplementary active compounds (e.g., preservatives,antibacterial, antiviral and antifungal agents) can also be incorporatedinto the compositions.

Cosolvents and adjuvants may be added to the formulation. Non-limitingexamples of cosolvents contain hydroxyl groups or other polar groups,for example, alcohols, such as isopropyl alcohol; glycols, such aspropylene glycol, polyethyleneglycol, polypropylene glycol, glycolether; glycerol; polyoxyethylene alcohols and polyoxyethylene fatty acidesters. Adjuvants include, for example, surfactants such as, soyalecithin and oleic acid; sorbitan esters such as sorbitan trioleate; andpolyvinylpyrrolidone.

A pharmaceutical composition contains a total amount of the activeingredient(s) sufficient to achieve an intended therapeutic effect.

The composition including a CSA may be administered with other activeagents. For example, in an embodiment, it may be administered with anantibiotic and/or antifungal agent. In an embodiment, the compositionmay be administered with an agent such as BACTIVATE, which serves toactivate dormant bacteria, causing the bacteria to quickly startdividing and multiplying, rendering the bacteria more easily treatablewith antibiotics and/or CSAs. BACTIVATE is available from University ofCopenhagen professor Anders Miki Bojesen and embryologist Morten RønnPetersen.

II. METHODS FOR ENHANCING FERTILITY IN A MAMMAL

Infectious causes of infertility (e.g., metritis, endometritis, oruterine infection) may be the most common cause of infertility in amammal, such as a mare. Such conditions often result in inflammation orinfection of the inner lining of the uterus, and are mostly commonlycaused by bacteria, although fungal infection also may occur.Endometritis affects up to 15% of broodmares. Endometritis is ofteninvisible, undetectable, and often under-diagnosed. The use of CSAsprovides a very broad spectrum non-toxic treatment that can be routinelyused by veterinarians or other medical practitioners, does not lead toresistance as do antibiotics, and is very economical. CSAs penetrate theepithelial tissue better than antibiotics, which results in a lowerincidence of recurrence than with treatments with antibiotics only. Inaddition, repeated use of antibiotics often leads to yeast and fungalinfections, which is not an issue with use of CSAs.

CSA compositions have been shown to be effective in the breakup ofbiofilm, and can even be used as a preventative treatment insub-clinical endometritis, where clinical signs are not apparent. Use ofa CSA lavage composition and/or infusion of a CSA composition provideexcellent treatment where uterine lavage may be recommended. In case ofcompostions formulated for lavage, the compositions may include fromabout 50 mg to about 200 mg of one or more CSA compounds in a suitablecarrier, such as saline solution, lactated ringer's solution, or sterilewater. Sometimes propylene glycol (e.g., 1%) can be added. For infusion,about 20 mg to about 100 mg of one or more CSA compounds can beadministered using any suitable carrier or delivery method. The infusionmay be typically used where an antibiotic infusion would be employed.

In addition to providing surprisingly improved results (e.g., allowingmares that could not successfully conceive and foal even with antibiotictreatment to do so), the presently described compositions and methodsprovide for improved retention of the CSAs in the uterus as compared toexisting alternatives. Of course, the CSA compositions and methods maybe employed in conjunction with antibiotics, antifungals, etc., with theadded advantage that the use of one or more CSA compounds reduces therequired or recommended amount of conventional antibiotics used foreffective treatment.

When fluid remains stagnant in the equine uterus, it is believed toweaken the effect of bacteria fighting white blood cells. It may flattenout the normal fold along the endometrial surface, which is lined withtiny hair-like cilia. When the folds are flattened out, it can becomemuch harder for the leukocytes to attack the bacteria, and the cilia maybecome far less effective in sweeping contaminants towards the cervix.Bacteria may be retained more easily on damaged areas of theendometrium, especially if the protective mucus blanket has been thinnedor eliminated due to persistent inflammation.

The mare's uterus is well-protected physically, with the cervix,vestibular-vaginal sphincter (hymen), and vulva lips all serving aseffective barriers against contaminants such as feces, urine, andbacteria. However, these contaminants can enter the uterus during matingor artificial insemination, as well as during estrus or veterinaryprocedures. To clear contaminants and dead sperm after breeding, theuterus sets off a natural inflammatory response. This acute form ofendometritis is a healthy, effective cleaning system that summons whiteblood cells (leukocytes) to attack and kill bacteria and rid the mare ofdead sperm. The debris is expelled from the uterus, which returns to itsnormal, uninflamed state. In healthy, fertile mares, this processtypically takes less than 2 days. Mares with a delayed inflammatoryresponse may not react immediately to the contaminants, however,allowing the contaminants to settle in the uterus, and in the case ofbacteria or fungi, to reproduce. The result can be increasedinflammation 3-4 days later, with secondary infection. This condition isreferred to as chronic or persistent endometritis, which causesinfertility.

In treating endometritis, many of the available antibiotics experienceresistance buildup, and in some cases, veterinarians use products thatare toxic. The CSA compositions do not experience resistance and are nottoxic.

According to some embodiments, a method for increasing fertility in amammal comprises: administering a therapeutically effective amount of atleast one cationic steroid antimicrobial (CSA) compound, or apharmaceutically acceptable salt thereof, to a reproductive structure ofa mammal. The reproductive structure can be a vagina, cervix, uterus,and combinations thereof in a female mammal or a penis in a male mammal(e.g., prior to copulation with a female).

The at least one CSA compound or pharmaceutically acceptable saltthereof can be administered to the reproductive structure of the mammalas a CSA-containing lavage and/or infusion composition including asolvent or liquid carrier and the at least one CSA compound orpharmaceutically acceptable salt thereof. Alternatively, theCSA-containing composition can be administered to the reproductivestructure of a mammal as an ointment or other topical compositionincluding a liquid or emollient carrier and the at least one CSA orpharmaceutically acceptable salt thereof. In some embodiments, both aCSA lavage (e.g., diluted with a large fraction of diluent) and a CSAinfusion (e.g., less dilute, less volume to be administered) may beapplied.

The CSA compound or pharmaceutically acceptable salt thereof killsmicrobes and/or breaks up microbial plaque or biofilm located on orwithin the reproductive structure. According to some embodiments, the atleast one CSA compound or pharmaceutically acceptable salt thereof killsat least 90% of one or more types of microbes. The at least one CSAcompound or pharmaceutically acceptable salt thereof may kill one ormore types of sperm-killing microbes where administered, while the atleast one CSA or pharmaceutically acceptable salt thereof and itsconcentration are adapted so that any residual CSA or pharmaceuticallyacceptable salt thereof that may remain on or in a treated reproductivestructure following treatment are adapted not to harm one or more typesof beneficial microbes typically present within the reproductivestructure of the mammal. Where any residual CSA or salt thereof remainson or in the reproductive structure, it may advantageously not poolwithin the uterus, but be dispersed.

The at least one CSA compound or pharmaceutically acceptable saltthereof can be present in a concentration sufficient to kill bothplanktonic and biofilm forms of microbes without causing harm tobeneficial microbes residing within the reproductive structure of themammal. The at least one CSA compound or pharmaceutically acceptablesalt thereof at least partially breaks up a microbial plaque or filmlocated within a uterus, which can cause of infertility in the mammal.

In some embodiments, the at least one CSA compound is delivered as partof a CSA-containing composition comprising a solvent or liquid carrier,at least one CSA compound, and a micelle-forming agent forming micellesencapsulating at least a portion of CSA molecules of the at least oneCSA compound so that no more than 25% of the CSA molecules formagglomerates larger than 1 micron in size.

According to one theory, the CSA-containing composition acts by breakingup biofilms and/or killing microbes that form biofilms present in theuterus or other reproductive structure of a mammal that might inhibitfertilization and/or implantation. Such biofilms may form a barrierbetween gametes (i.e., sperm cells and the ovum) and/or preventimplantation of a fertilized zygote within the uterus and/or causespontaneous abortion of a zygote shortly after implantation. It isbelieved that CSA-containing compositions are able to break up biofilmsand/or kill microbes that form such biofilms as a result of the uniqueamphiphilic nature and chemical functionalities contained in CSAcompounds. The CSA-containing composition can hasten uterine epithelialcell healing due to its epitheliotrophic activity and modulates theinflammation due to enhanced innate immune response. Use ofCSA-containing compositions as described herein has been found toeffectively treat endometritis and inflammation associated therewith,allowing the female to conceive and have a successful pregnancy.

Compositions that include CSA compounds may be formulated to beespecially beneficial for enhancing fertility on mammals. In someembodiments, the CSA-containing compositions can be formulated in orderto deliver CSA compounds as individually dispersed CSA molecules and/orvery small CSA particles having low agglomeration. According to oneembodiment, CSA particles are formulated in order to remain stable andresist agglomeration to larger CSA particles over a desired time period.This may be accomplished, for example, by the use of micelle-formingagents that are able to form micelles in a liquid carrier, whichencapsulate the CSA molecules and prevent or reduce agglomeration.

III. EXAMPLES Example 1

A study was undertaken to determine the histologic effect that infusionof the present lavages into the uterus for four consecutive days wouldhave on the endometrium, and to determine the pregnancy rates followinga single infusion of a CSA-containing lavage 24 hours post breeding.

In part one of the study protocol, the histologic effect ofintra-uterine infusion of the composition on the endometrium wasevaluated. Part 1 of the protocol was as follows:

-   -   Day 0: culture, cytology, and first biopsy obtained when mare        has a 25 mm follicle, moderately relaxed cervix, and greater        than or equal to 1.5 uterine edema.    -   Day 1: infuse 50 mL of sterile saline and 10 mL of a composition        comprising 20 mg to 100 mg CSA (“Ceragyn CSA”) (e.g., 50 mg        CSA-44).    -   Day 2: lavage with 1 L Lactated Ringer's Solution (LRS),        followed by infusion of 50 mL sterile saline and 10 mL Ceragyn        CSA.    -   Day 3: lavage with 1 L LRS, followed by infusion of 50 mL        sterile saline and 10 mL Ceragyn CSA.    -   Day 4: lavage with 1 L LRS, followed by infusion of 50 mL        sterile saline and 10 mL Ceragyn CSA.    -   Day 5: lavage with 1 L LRS.    -   Day 6: biopsy.    -   Next estrus cycle, culture, cytology, and biopsy obtained when        mare has 25 mm follicle, moderately relaxed cervix and greater        than or equal to 1.5 uterine edema.

The results of part 1 of the protocol showed no significant differencein endometrial biopsy scores prior to and post intra-uterine infusion ofCeragyn CSA. That means that Ceragyn CSA had no measurable negativehistologic effect.

Part 2 of the protocol was directed to determining pregnancy ratesfollowing a single intra-uterine infusion of Ceragyn CSA post breeding.Part 2 of the protocol was as follows:

-   -   Day 0: 2,500 IU of hCG was administered intravenously when there        was a 35 mm follicle and grade 3 uterine edema.    -   Day 1: inseminate 1 billion sperm (>30% progressive motility).    -   Day 2: infused 60 cc solution containing 10 cc Ceragyn CSA        diluted in 50 cc saline.    -   Day 3: mares were ultrasounded per rectum daily until ovulation        was detected and evaluated for pregnancy 14 days post ovulation.

For the mares that fulfilled these criteria (5 mares, 6 cycles), 4cycles resulted in pregnancy and 2 cycles did not (66% per cycleconception rate). Of the 2 mares that did not conceive, with 1 mare theexperiment was repeated and she became pregnant. The other mare did notbecome pregnant when subsequently bred without Ceragyn CSA.

Example 2

A study was undertaken to evaluate use of a CSA-containing composition(Ceragyn CSA) in mares. The facility where the study was undertakenspecializes in embryo transfers and frozen embryos. The mares treated inthis example had previously been at a leading school of veterinarymedicine for a period of two years, and during that time, theveterinarians in charge of treatment were unable to extract a viableembryo due to yeast infections and other complications.

When the mares arrived at the facility where Ceragyn CSA was beingevaluated, they were first treated with conventional antibiotics.Following conventional treatment with antibiotics, the attendingveterinarians were not able to extract any viable embryo from the mares.

After learning about Ceragyn CSA, the attending veterinarians usedCeragyn CSA in the two mares, along with other recipient mares that hadstruggled to retain a pregnancy. Treatment with Ceragyn CSA includedapplication of a Ceragyn CSA uterine lavage providing about 50 mg toabout 200 mg CSA (e.g., 100 mg CSA-44) to flush the uterus up to 4 hoursprior to insemination and/or 6 to 48 hours after breeding. Lavagetreatment prior to insemination is helpful in treating infertilityassociated with fluid retained in the uterus. Lavage treatmentpost-breeding is helpful in treating post breeding endometriosis.

For each lavage treatment, 60 mL of the Ceragyn CSA composition wasmixed with 1 L sterile water, saline, or Lactated Ringer's Solution(LRS) and administered using a sterile catheter. Treatment with CeragynCSA also included application of a Ceragyn CSA uterine infusion to treatgram-negative and gram-positive infections, as well as yeast infections.For each infusion treatment, 60 mL of the Ceragyn CSA composition wastransferred into a sterile syringe and aseptically administered into theuterus using a sterile catheter or pipette. After such treatment, theattending veterinarians were able to successfully complete an embryotransfer.

Example 3

A top quality veterinary medical and surgical services facility had a 16year old Quarter Horse mare that had been unable to get in foal for twoyears. Treatment proceeded as follows. Initially, a 1 cm by 3 cm tissuesection was submitted from the endometrium of the mare to AntechDiagnostics, in Irvine, Calif. This section of endometrium was describedby Antech as containing a diffuse infiltration of mixed inflammatorycells embedded within the stratum compactum and accompanied by diffuseedema. The overlying mucosal epithelium was intact. The mixedinflammatory reaction was composed of lymphocytes, neutrophils, andplasma cells. Rare foci of inflammatory cell exocytosis were also noted.The supporting stratum spongiosum exhibited diffuse edema. Nosignificant glandular nesting, clumping, or periglandular fibrosis wereidentified.

The microscopic findings from the lab were described as mild to moderatediffuse acute superficial endometriosis, Grade IIA endometrium. The labalso isolated Escherichia coli and Alpha-hemolytic streptococci. Thesensitivity panel yielded the results in Table 1.

TABLE 1 Sensitivity Panel Alpha hemolytic Escherichia coli streptococciAntibiotic Organism Isolated organism isolated Amikacin SensitiveResistant Ampicillin Intermediate Sensitive Cefazolin SensitiveSensitive Ceftiofur Sensitive Sensitive Enrofloxacin Sensitive SensitiveErythromycin N/A Sensitive Florfenicol Sensitive N/A GentamicinSensitive Resistant Oxytetracycline Resistant Sensitive Rifampin N/ASensitive Ticarcillin Sensitive Sensitive Trimmicosin Intermediate N/ATMP/Sulfa Resistant Sensitive

Twenty-one (21) days later, when the mare was coming back in heat, themare was lavaged with 2 mL Amikacin and 60 mL of Ceragyn CSA uterinelavage combined in a liter of normal saline. On the next day, the uterushad pooled fluid of approximately 250 mL. The prior lavage treatment wasrepeated along with 40 IU oxytocin immediately after lavage and repeatedin 4 hour intervals for 3 successive treatments. At the time of thelavage, the mare was given 1 mL Deslorelin. One day later, the mare hadno fluid in uterus and was taken to be bred live cover. The ownerconfirmed that the mare foaled approximately 11 months later. In theattending veterinarian's opinion, the Ceragyn CSA treatment made thedifference as other treatments had developed resistance.

The present invention may be embodied in other specific forms withoutdeparting from its spirit or essential characteristics. The describedembodiments are to be considered in all respects only as illustrativeand not restrictive. The scope of the invention is, therefore, indicatedby the appended claims rather than by the foregoing description. Allchanges which come within the meaning and range of equivalency of theclaims are to be embraced within their scope.

What is claimed is:
 1. A method for increasing fertility in a mammal,comprising: administering a therapeutically effective amount of at leastone cationic steroid antimicrobial (CSA) compound of Formula (I), or apharmaceutically acceptable salt thereof, to a reproductive structure ofa mammal:

where, rings A, B, C, and D are independently saturated, or are fully orpartially unsaturated, provided that at least two of rings A, B, C, andD are saturated; m, n, p, and q are independently 0 or 1; R₁ through R₄,R₆, R₇, R₁₁, R₁₂, R₁₅, R₁₆, and R₁₈ are independently selected from thegroup consisting of hydrogen, hydroxyl, a substituted or unsubstitutedalkyl, a substituted or unsubstituted hydroxyalkyl, a substituted orunsubstituted alkyloxyalkyl, a substituted or unsubstitutedalkylcarboxyalkyl, a substituted or unsubstituted alkylaminoalkyl, asubstituted or unsubstituted alkylaminoalkylamino, a substituted orunsubstituted alkylaminoalkylamino-alkylamino, a substituted orunsubstituted aminoalkyl, a substituted or unsubstituted aryl, asubstituted or unsubstituted arylaminoalkyl, a substituted orunsubstituted haloalkyl, a substituted or unsubstituted alkenyl, asubstituted or unsubstituted alkynyl, oxo, a linking group attached to asecond steroid, a substituted or unsubstituted aminoalkyloxy, asubstituted or unsubstituted aminoalkyloxyalkyl, a substituted orunsubstituted aminoalkylcarboxy, a substituted or unsubstitutedaminoalkylaminocarbonyl, a substituted or unsubstitutedaminoalkylcarboxamido, a substituted or unsubstituteddi(alkyl)aminoalkyl, H₂N—HC(Q₅)-C(O)—O—, H₂N—HC(Q₅)-C(O)—N(H)—, asubstituted or unsubstituted azidoalkyloxy, a substituted orunsubstituted cyanoalkyloxy, P.G.-HN—HC(Q₅)-C(O)—O—, a substituted orunsubstituted guanidinoalkyloxy, a substituted or unsubstitutedquaternary ammonium alkylcarboxy, and a substituted or unsubstitutedguanidinoalkyl carboxy, where Q₅ is a side chain of an amino acid andP.G. is an amino protecting group; and R₅, R₈, R₉, R₁₀, R₁₃, R₁₄ and R₁₇are independently deleted when one of rings A, B, C, or D is unsaturatedso as to complete the valency of the carbon atom at that site, or whenpresent, R₅, R₈, R₉, R₁₀, R₁₃, and R₁₄ are independently selected fromthe group consisting of hydrogen, hydroxyl, a substituted orunsubstituted alkyl, a substituted or unsubstituted hydroxyalkyl, asubstituted or unsubstituted alkyloxyalkyl, a substituted orunsubstituted aminoalkyl, a substituted or unsubstituted aryl, asubstituted or unsubstituted haloalkyl, a substituted or unsubstitutedalkenyl, a substituted or unsubstituted alkynyl, oxo, a linking groupattached to a second steroid, a substituted or unsubstitutedaminoalkyloxy, a substituted or unsubstituted aminoalkylcarboxy, asubstituted or unsubstituted aminoalkylaminocarbonyl, a substituted orunsubstituted di(alkyl)aminoalkyl, H₂N—HC(Q₅)-C(O)—O—,H₂N—HC(Q₅)-C(O)—N(H)—, azidoalkyloxy, cyano-alkyloxy,P.G.-HN—HC(Q₅)-C(O)—O—, guanidinoalkyloxy, and guanidine-alkylcarboxy,where Q₅ is a side chain of an amino acid and P.G. is an aminoprotecting group, provided that at least two or three of R₁₋₄, R₆, R₇,R₁₁, R₁₂, R₁₅, R₁₆, R₁₇, and R₁₈ are independently selected from thegroup consisting of a substituted or unsubstituted aminoalkyl, asubstituted or unsubstituted aminoalkyloxy, a substituted orunsubstituted alkylcarboxyalkyl, a substituted or unsubstitutedalkylaminoalkylamino, a substituted or unsubstitutedalkylaminoalkylamino-alkylamino, a substituted or unsubstitutedaminoalkylcarboxy, a substituted or unsubstituted arylaminoalkyl, asubstituted or unsubstituted aminoalkyloxyaminoalkylaminocarbonyl, asubstituted or unsubstituted aminoalkylamino-carbonyl, a substituted orunsubstituted aminoalkylcarboxyamido, a quaternaryammoniumalkylcarboxy,a substituted or unsubstituted di(alkyl)aminoalkyl, H₂N—HC(Q₅)-C(O)—O—,H₂N—HC(Q₅)-C(O)—N(H)—, azidoalkyloxy, cyanoalkyloxy,P.G.-HN—HC(Q₅)-C(O)—O—, a substituted or unsubstitutedguanidinoalkyloxy, and a substituted or unsubstitutedguanidinoalkylcarboxy, and the administered at least one CSA compound,or pharmaceutically acceptable salt thereof, at least partially breakingup a bacterial biofilm associated with the reproductive structure of themammal to increase fertility of the mammal.
 2. A method as in claim 1,wherein the reproductive structure of the mammal is selected from thegroup consisting of a vagina, cervix, uterus, and combinations thereofof a female mammal.
 3. A method as in claim 1, wherein the at least oneCSA compound or pharmaceutically acceptable salt thereof is administeredto the reproductive structure of the mammal as a lavage and/or infusioncomposition including a solvent or liquid carrier and the at least oneCSA compound or pharmaceutically acceptable salt thereof.
 4. A method asin claim 1, wherein the reproductive structure of the mammal is a penisof a male mammal.
 5. A method as in claim 1, wherein the at least oneCSA compound or pharmaceutically acceptable salt thereof is administeredto the reproductive structure of the mammal as an ointment or othertopical composition including a liquid or emollient carrier and the atleast one CSA compound or pharmaceutically acceptable salt thereof.
 6. Amethod as in claim 1, wherein at least one bacterial strain associatedwith the bacterial biofilm interferes with sperm activity, and whereinthe administered at least one CSA compound or pharmaceuticallyacceptable salt thereof kills the at least one bacterial strain thatinterferes with sperm activity.
 7. A method as in claim 1, wherein theat least one CSA compound or pharmaceutically acceptable salt thereofkills at least 90% of one or more types of microbes.
 8. A method as inclaim 1, wherein infertility is caused by metritis or endometritis andwherein the at least one CSA compound or pharmaceutically acceptablesalt thereof hastens uterine epithelial cell healing and modulatesinflammation.
 9. A method as in claim 1, wherein the at least one CSAcompound or pharmaceutically acceptable salt thereof is adapted to andpresent in a concentration sufficient to kill both planktonic andbiofilm forms of sperm killing microbes without causing harm tobeneficial microbes residing within the reproductive structure of themammal.
 10. A method as in claim 1, wherein the mammal is a non-humanmammal.
 11. A method as in claim 10, wherein the non-human mammal is ahorse, bovine, pig, dog, or cat.
 12. A method as in claim 1, wherein themammal is a human.
 13. A method as in claim 1, wherein the CSA compoundcomprises a plurality of cationic groups attached to a steroidalbackbone, the cationic groups being attached to the steroidal backbonethrough hydrolysable ester linkages.
 14. A method as in claim 1, whereinthe at least one CSA compound or pharmaceutically acceptable saltthereof is administered using a CSA-containing composition comprising: asolvent or liquid carrier; the at least one CSA compound orpharmaceutically acceptable salt thereof; and a micelle-forming agentforming micelles encapsulating at least a portion of CSA molecules ofthe at least one CSA compound or pharmaceutically acceptable saltthereof so that no more than 25% of the CSA molecules form agglomerateslarger than 1 micron in size.
 15. A method as in claim 1, wherein: R₁,R₂, R₄, R₅, R₆, R₈, R₁₀, R₁₁, R₁₄, R₁₆, and R₁₇ are each hydrogen; R₉and R₁₃ are each methyl; and R₃, R₇, R₁₂, and R₁₈ are independentlyselected from the group consisting of aminoalkyloxy; aminoalkylcarboxy;alkylaminoalkyl; alkoxycarbonylalkyl; alkyl-carbonyl alkyl;di(alkyl)aminoalkyl; alkoxycarbonylalkyl; and alkylcarboxyalkyl.
 16. Amethod as in claim 1, wherein the at least one CSA compound orpharmaceutically acceptable salt thereof is selected from the groupconsisting of:

and pharmaceutically acceptable salts thereof.
 17. A method as in claim1, wherein the at least one CSA compound or pharmaceutically acceptablesalt thereof comprises:

or pharmaceutically acceptable salt thereof.
 18. A method as in claim 1,wherein the pharmaceutically acceptable salt is a hydrochloride salt ora tri-hydrochloride salt.
 19. A method for increasing fertility in amammal having metritis or endometritis, comprising: administering atherapeutically effective amount of at least one cationic steroidantimicrobial (CSA) compound, or a pharmaceutically acceptable saltthereof, to a reproductive structure of the mammal, wherein theadministered at least one CSA compound, or pharmaceutically acceptablesalt thereof, at least partially breaks up a biofilm associated with thereproductive structure of the mammal and/or hastens uterine epithelialcell healing and modulates inflammation to increase fertility in themammal, and wherein the at least one CSA compound is a compound ofFormula (IA) or a pharmaceutically acceptable salt thereof:

where, rings A, B, C, and D are independently saturated; R₁ through R₄,R₆, R₇, R₁₁, R₁₂, R₁₆, and R₁₈ are independently selected from the groupconsisting of hydrogen, hydroxyl, a substituted or unsubstituted alkyl,a substituted or unsubstituted hydroxyalkyl, a substituted orunsubstituted alkyloxyalkyl, a substituted or unsubstitutedalkylcarboxyalkyl, a substituted or unsubstituted alkylaminoalkyl, asubstituted or unsubstituted alkylamino-alkylamino, a substituted orunsubstituted alkylaminoalkylamino-alkylamino, a substituted orunsubstituted aminoalkyl, a substituted or unsubstituted aryl, asubstituted or unsubstituted arylaminoalkyl, a substituted orunsubstituted haloalkyl, a substituted or unsubstituted alkenyl, asubstituted or unsubstituted alkynyl, oxo, a linking group attached to asecond steroid, a substituted or unsubstituted aminoalkyloxy, asubstituted or unsubstituted aminoalkyloxyalkyl, a substituted orunsubstituted aminoalkylcarboxy, a substituted or unsubstitutedaminoalkylaminocarbonyl, a substituted or unsubstitutedaminoalkylcarboxamido, a substituted or unsubstituteddi(alkyl)aminoalkyl, H₂N—HC(Q₅)-C(O)—O—, H₂N—HC(Q₅)-C(O)—N(H)—, asubstituted or unsubstituted azido-alkyloxy, a substituted orunsubstituted cyanoalkyloxy, P.G.-HN—HC(Q₅)-C(O)—O—, a substituted orunsubstituted guanidinoalkyloxy, a substituted or unsubstitutedquaternary ammonium alkylcarboxy, and a substituted or unsubstitutedguanidinoalkyl carboxy, where Q₅ is a side chain of an amino acid andP.G. is an amino protecting group; and R₅, R₈, R₉, R₁₀, R₁₃, and R₁₄ areindependently selected from the group consisting of hydrogen, hydroxyl,a substituted or unsubstituted alkyl, a substituted or unsubstitutedhydroxyalkyl, a substituted or unsubstituted alkyloxyalkyl, asubstituted or unsubstituted aminoalkyl, a substituted or unsubstitutedaryl, a substituted or unsubstituted haloalkyl, a substituted orunsubstituted alkenyl, a substituted or unsubstituted alkynyl, oxo, alinking group attached to a second steroid, a substituted orunsubstituted aminoalkyloxy, a substituted or unsubstitutedaminoalkylcarboxy, a substituted or unsubstitutedaminoalkylaminocarbonyl, a substituted or unsubstituteddi(alkyl)aminoalkyl, H₂N—HC(Q₅)-C(O)—O—, H₂N—HC(Q₅)-C(O)—N(H)—,azidoalkyloxy, cyano-alkyloxy, P.G.-HN—HC(Q₅)-C(O)—O—,guanidinoalkyloxy, and guanidine-alkylcarboxy, where Q₅ is a side chainof an amino acid and P.G. is an amino protecting group, provided that atleast two or three of R₁₋₄, R₆, R₇, R₁₁, R₁₂, R₁₆, R₁₇, and R₁₈ areindependently selected from the group consisting of a substituted orunsubstituted aminoalkyl, a substituted or unsubstituted aminoalkyloxy,a substituted or unsubstituted alkylcarboxyalkyl, a substituted orunsubstituted alkylaminoalkylamino, a substituted or unsubstitutedalkylaminoalkylamino-alkylamino, a substituted or unsubstitutedaminoalkylcarboxy, a substituted or unsubstituted arylaminoalkyl, asubstituted or unsubstituted aminoalkyloxyaminoalkylaminocarbonyl, asubstituted or unsubstituted aminoalkylamino-carbonyl, a substituted orunsubstituted aminoalkylcarboxyamido, a quaternaryammoniumalkylcarboxy,a substituted or unsubstituted di(alkyl)aminoalkyl, H₂N—HC(Q₅)-C(O)—O—,H₂N—HC(Q₅)-C(O)—N(H)—, azidoalkyloxy, cyanoalkyloxy,P.G.-HN—HC(Q₅)-C(O)—O—, a substituted or unsubstitutedguanidinoalkyloxy, and a substituted or unsubstitutedguanidinoalkylcarboxy.
 20. A method as in claim 19, wherein: R₁, R₂, R₄,R₅, R₆, R₈, R₁₀, R₁₁, R₁₄, R₁₆, and R₁₇ are each hydrogen; R₉ and R₁₃are each methyl; and R₃, R₇, R₁₂, and R₁₈ are independently selectedfrom the group consisting of aminoalkyloxy; aminoalkylcarboxy;alkylaminoalkyl; alkoxycarbonylalkyl; alkylcarbonylalkyl;di(alkyl)aminoalkyl; alkoxycarbonylalkyl; and alkyl carboxyalkyl.
 21. Amethod as in claim 19, wherein: R₃, R₇, R₁₂, and R₁₈ are independentlyselected from the group consisting of hydrogen, an unsubstituted(C₁-C₁₈) alkyl, unsubstituted (C₁-C₁₈) hydroxyalkyl, unsubstituted(C₁-C₁₈) alkyloxy-(C₁-C₁₈) alkyl, unsubstituted (C₁-C₁₈)alkylcarboxy-(C₁-C₁₈) alkyl, unsubstituted (C₁-C₁₈)alkylamino-(C₁-C₁₈)alkyl, unsubstituted (C₁-C₁₈) alkylamino-(C₁-C₁₈)alkylamino, unsubstituted (C₁-C₁₈) alkylamino-(C₁-C₁₈)alkylamino-(C₁-C₁₈) alkylamino, an unsubstituted (C₁-C₁₈) aminoalkyl, anunsubstituted arylamino-(C₁-C₁₈) alkyl, an unsubstituted (C₁-C₁₈)aminoalkyloxy, an unsubstituted (C₁-C₁₈) aminoalkyloxy-(C₁-C₁₈) alkyl,an unsubstituted (C₁-C₁₈) aminoalkylcarboxy, an unsubstituted (C₁-C₁₈)aminoalkylaminocarbonyl, an unsubstituted (C₁-C₁₈)aminoalkylcarboxamido, an unsubstituted di(C₁-C₁₈ alkyl)aminoalkyl,unsubstituted (C₁-C₁₈) guanidinoalkyloxy, unsubstituted (C₁-C₁₈)quaternaryammoniumalkylcarboxy, and unsubstituted (C₁-C₁₈)guanidinoalkylcarboxy; and R₁, R₂, R₄, R₅, R₆, R₈, R₉, R₁₀, R₁₁, R₁₃,R₁₄, R₁₆, and R₁₇ are independently selected from the group consistingof hydrogen and unsubstituted (C₁-C₆) alkyl.
 22. A method as in claim19, wherein: R₃, R₇, R₁₂, and R₁₈ are independently selected from thegroup consisting of hydrogen, an unsubstituted (C₁-C₆) alkyl,unsubstituted (C₁-C₆) hydroxyalkyl, unsubstituted (C₁-C₁₆)alkyloxy-(C₁-C₅) alkyl, unsubstituted (C₁-C₁₆) alkylcarboxy-(C₁-C₅)alkyl, unsubstituted (C₁-C₁₆) alkylamino-(C₁-C₅)alkyl, (C₁-C₁₆)alkylamino-(C₁-C₅) alkylamino, unsubstituted (C₁-C₁₆)alkylamino-(C₁-C₁₆) alkylamino-(C₁-C₅) alkylamino, an unsubstituted(C₁-C₁₆) aminoalkyl, an unsubstituted arylamino-(C₁-C₅) alkyl, anunsubstituted (C₁-C₅) aminoalkyloxy, an unsubstituted (C₁-C₁₆)aminoalkyloxy-(C₁-C₅) alkyl, an unsubstituted (C₁-C₅) aminoalkylcarboxy,an unsubstituted (C₁-C₅) aminoalkylaminocarbonyl, an unsubstituted(C₁-C₅) aminoalkylcarboxamido, an unsubstituted di(C₁-C₅alkyl)amino-(C₁-C₅) alkyl, unsubstituted (C₁-C₅) guanidinoalkyloxy,unsubstituted (C₁-C₁₆) quaternaryammoniumalkylcarboxy, and unsubstituted(C₁-C₁₆) guanidinoalkylcarboxy; R₁, R₂, R₄, R₅, R₆, R₈, R₁₀, R₁₁, R₁₄,R₁₆, and R₁₇ are each hydrogen; R₉ and R₁₃ are each methyl; and R₃, R₇,R₁₂, and R₁₈ are independently selected from the group consisting ofaminoalkyloxy; aminoalkylcarboxy; alkylaminoalkyl; alkoxycarbonylalkyl;alkyl-carbonyl alkyl; di(alkyl)aminoalkyl; alkoxycarbonylalkyl; andalkyl carboxyalkyl.
 23. A method as in claim 19, wherein R₃, R₇, and R₁₂are the same and are selected from the group consisting of aminoalkyloxyand aminoalkylcarboxy; and R₁₈ is selected from the group consisting ofalkylaminoalkyl; alkoxycarbonylalkyl; alkylcarbonyloxyalkyl;di(alkyl)aminoalkyl; C-carboxyalkyl; alkylaminoalkyl;alkyoxycarbonylalkyl; and alkylcarboxyalkyl.
 24. A method as in claim19, wherein R₃, R₇, R₁₂, and R₁₈ are independently selected from thegroup consisting of amino-C₃-alkyloxy; amino-C₃-alkyl-carboxy;C₅-alkylamino-C₅-alkyl; C₈-alkoxy-carbonyl-C₄-alkyl;C₁₀-alkoxy-carbonyl-C₄-alkyl; C₈-alkyl-carbonyl-C₄-alkyl;di-(C₅-alkyl)amino-C₅-alkyl; C₁₃-alkylamino-C₅-alkyl;C₆-alkoxy-carbonyl-C₄-alkyl; C₆-alkyl-carboxy-C₄-alkyl; andC₁₆-alkylamino-C₅-alkyl.
 25. A method for increasing fertility in ahorse, comprising: administering a therapeutically effective amount ofat least one cationic steroid antimicrobial (CSA) compound of Formula(I), or a pharmaceutically acceptable salt thereof, to a reproductivestructure of the horse in need thereof:

where, rings A, B, C, and D are independently saturated, or are fully orpartially unsaturated, provided that at least two of rings A, B, C, andD are saturated; m, n, p, and q are independently 0 or 1; R₁ through R₄,R₆, R₇, R₁₁, R₁₂, R₁₅, R₁₆, and R₁₈ are independently selected from thegroup consisting of hydrogen, hydroxyl, a substituted or unsubstitutedalkyl, a substituted or unsubstituted hydroxyalkyl, a substituted orunsubstituted alkyloxyalkyl, a substituted or unsubstitutedalkylcarboxyalkyl, a substituted or unsubstituted alkylaminoalkyl, asubstituted or unsubstituted alkylaminoalkylamino, a substituted orunsubstituted alkylaminoalkylamino-alkylamino, a substituted orunsubstituted aminoalkyl, a substituted or unsubstituted aryl, asubstituted or unsubstituted arylaminoalkyl, a substituted orunsubstituted haloalkyl, a substituted or unsubstituted alkenyl, asubstituted or unsubstituted alkynyl, oxo, a linking group attached to asecond steroid, a substituted or unsubstituted aminoalkyloxy, asubstituted or unsubstituted aminoalkyloxyalkyl, a substituted orunsubstituted aminoalkylcarboxy, a substituted or unsubstitutedaminoalkylaminocarbonyl, a substituted or unsubstitutedaminoalkylcarboxamido, a substituted or unsubstituteddi(alkyl)aminoalkyl, H₂N—HC(Q₅)-C(O)—O—, H₂N—HC(Q₅)-C(O)—N(H)—, asubstituted or unsubstituted azidoalkyloxy, a substituted orunsubstituted cyanoalkyloxy, P.G.-HN—HC(Q₅)-C(O)—O—, a substituted orunsubstituted guanidinoalkyloxy, a substituted or unsubstitutedquaternary ammonium alkylcarboxy, and a substituted or unsubstitutedguanidinoalkyl carboxy, where Q₅ is a side chain of an amino acid andP.G. is an amino protecting group; and R₅, R₈, R₉, R₁₀, R₁₃, R₁₄ and R₁₇are independently deleted when one of rings A, B, C, or D is unsaturatedso as to complete the valency of the carbon atom at that site, or whenpresent, R₅, R₈, R₉, R₁₀, R₁₃, and R₁₄ are independently selected fromthe group consisting of hydrogen, hydroxyl, a substituted orunsubstituted alkyl, a substituted or unsubstituted hydroxyalkyl, asubstituted or unsubstituted alkyloxyalkyl, a substituted orunsubstituted aminoalkyl, a substituted or unsubstituted aryl, asubstituted or unsubstituted haloalkyl, a substituted or unsubstitutedalkenyl, a substituted or unsubstituted alkynyl, oxo, a linking groupattached to a second steroid, a substituted or unsubstitutedaminoalkyloxy, a substituted or unsubstituted aminoalkylcarboxy, asubstituted or unsubstituted aminoalkylaminocarbonyl, a substituted orunsubstituted di(alkyl)aminoalkyl, H₂N—HC(Q₅)-C(O)—O—,H₂N—HC(Q₅)-C(O)—N(H)—, azidoalkyloxy, cyano-alkyloxy,P.G.-HN—HC(Q₅)-C(O)—O—, guanidinoalkyloxy, and guanidine-alkylcarboxy,where Q₅ is a side chain of an amino acid and P.G. is an aminoprotecting group, provided that at least two or three of R₁₋₄, R₆, R₇,R₁₁, R₁₂, R₁₅, R₁₆, R₁₇, and R₁₈ are independently selected from thegroup consisting of a substituted or unsubstituted aminoalkyl, asubstituted or unsubstituted aminoalkyloxy, a substituted orunsubstituted alkylcarboxyalkyl, a substituted or unsubstitutedalkylaminoalkylamino, a substituted or unsubstitutedalkylaminoalkylamino-alkylamino, a substituted or unsubstitutedaminoalkylcarboxy, a substituted or unsubstituted arylaminoalkyl, asubstituted or unsubstituted aminoalkyloxyaminoalkylaminocarbonyl, asubstituted or unsubstituted aminoalkylamino-carbonyl, a substituted orunsubstituted aminoalkylcarboxyamido, a quaternaryammoniumalkylcarboxy,a substituted or unsubstituted di(alkyl)aminoalkyl, H₂N—HC(Q₅)-C(O)—O—,H₂N—HC(Q₅)-C(O)—N(H)—, azidoalkyloxy, cyanoalkyloxy,P.G.-HN—HC(Q₅)-C(O)—O—, a substituted or unsubstitutedguanidinoalkyloxy, and a substituted or unsubstitutedguanidinoalkylcarboxy, and the at least one CSA compound increasingfertility of the horse.
 26. The method as in claim 25, wherein theadministered at least one CSA compound, or pharmaceutically acceptablesalt thereof, at least partially breaks up a bacterial biofilmassociated with the reproductive structure of the horse to increasefertility of the horse.
 27. A method as in claim 25, wherein the atleast one CSA compound or pharmaceutically acceptable salt thereof isselected from the group consisting of:

and pharmaceutically acceptable salts thereof.
 28. The method as inclaim 25, wherein infertility is caused by metritis or endometritis, andwherein the at least one CSA compound, or pharmaceutically acceptablesalt thereof, hastens uterine epithelial cell healing and modulatesinflammation.
 29. A method for increasing fertility in a mammal,comprising: administering a lavage composition comprising a solvent orliquid carrier and a therapeutically effective amount of at least onecationic steroid antimicrobial (CSA) compound of Formula (I), or apharmaceutically acceptable salt thereof, to a reproductive structure ofa mammal in order to increase fertility of the mammal:

where, rings A, B, C, and D are independently saturated, or are fully orpartially unsaturated, provided that at least two of rings A, B, C, andD are saturated; m, n, p, and q are independently 0 or 1; R₁ through R₄,R₆, R₇, R₁₁, R₁₂, R₁₅, R₁₆, and R₁₈ are independently selected from thegroup consisting of hydrogen, hydroxyl, a substituted or unsubstitutedalkyl, a substituted or unsubstituted hydroxyalkyl, a substituted orunsubstituted alkyloxyalkyl, a substituted or unsubstitutedalkylcarboxyalkyl, a substituted or unsubstituted alkylaminoalkyl, asubstituted or unsubstituted alkylaminoalkylamino, a substituted orunsubstituted alkylaminoalkylamino-alkylamino, a substituted orunsubstituted aminoalkyl, a substituted or unsubstituted aryl, asubstituted or unsubstituted arylaminoalkyl, a substituted orunsubstituted haloalkyl, a substituted or unsubstituted alkenyl, asubstituted or unsubstituted alkynyl, oxo, a linking group attached to asecond steroid, a substituted or unsubstituted aminoalkyloxy, asubstituted or unsubstituted aminoalkyloxyalkyl, a substituted orunsubstituted aminoalkylcarboxy, a substituted or unsubstitutedaminoalkylaminocarbonyl, a substituted or unsubstitutedaminoalkylcarboxamido, a substituted or unsubstituteddi(alkyl)aminoalkyl, H₂N—HC(Q₅)-C(O)—O—, H₂N—HC(Q₅)-C(O)—N(H)—, asubstituted or unsubstituted azidoalkyloxy, a substituted orunsubstituted cyanoalkyloxy, P.G.-HN—HC(Q₅)-C(O)—O—, a substituted orunsubstituted guanidinoalkyloxy, a substituted or unsubstitutedquaternary ammonium alkylcarboxy, and a substituted or unsubstitutedguanidinoalkyl carboxy, where Q₅ is a side chain of an amino acid andP.G. is an amino protecting group; and R₅, R₈, R₉, R₁₀, R₁₃, R₁₄ and R₁₇are independently deleted when one of rings A, B, C, or D is unsaturatedso as to complete the valency of the carbon atom at that site, or whenpresent, R₅, R₈, R₉, R₁₀, R₁₃, and R₁₄ are independently selected fromthe group consisting of hydrogen, hydroxyl, a substituted orunsubstituted alkyl, a substituted or unsubstituted hydroxyalkyl, asubstituted or unsubstituted alkyloxyalkyl, a substituted orunsubstituted aminoalkyl, a substituted or unsubstituted aryl, asubstituted or unsubstituted haloalkyl, a substituted or unsubstitutedalkenyl, a substituted or unsubstituted alkynyl, oxo, a linking groupattached to a second steroid, a substituted or unsubstitutedaminoalkyloxy, a substituted or unsubstituted aminoalkylcarboxy, asubstituted or unsubstituted aminoalkylaminocarbonyl, a substituted orunsubstituted di(alkyl)aminoalkyl, H₂N—HC(Q₅)-C(O)—O—,H₂N—HC(Q₅)-C(O)—N(H)—, azidoalkyloxy, cyano-alkyloxy,P.G.-HN—HC(Q₅)-C(O)—O—, guanidinoalkyloxy, and guanidine-alkylcarboxy,where Q₅ is a side chain of an amino acid and P.G. is an aminoprotecting group, provided that at least two or three of R₁₋₄, R₆, R₇,R₁₁, R₁₂, R₁₅, R₁₆, R₁₇, and R₁₈ are independently selected from thegroup consisting of a substituted or unsubstituted aminoalkyl, asubstituted or unsubstituted aminoalkyloxy, a substituted orunsubstituted alkylcarboxyalkyl, a substituted or unsubstitutedalkylaminoalkylamino, a substituted or unsubstitutedalkylaminoalkylamino-alkylamino, a substituted or unsubstitutedaminoalkylcarboxy, a substituted or unsubstituted arylaminoalkyl, asubstituted or unsubstituted aminoalkyloxy-aminoalkylaminocarbonyl, asubstituted or unsubstituted aminoalkylamino-carbonyl, a substituted orunsubstituted aminoalkylcarboxyamido, a quaternaryammoniumalkylcarboxy,a substituted or unsubstituted di(alkyl)aminoalkyl, H₂N—HC(Q₅)-C(O)—O—,H₂N—HC(Q₅)-C(O)—N(H)—, azidoalkyloxy, cyanoalkyloxy,P.G.-HN—HC(Q₅)-C(O)—O—, a substituted or unsubstitutedguanidinoalkyloxy, and a substituted or unsubstitutedguanidinoalkylcarboxy.